Fresh blastocyst (day 5 to 6) stage transfer seems to be more effective than cleavage (day 2 or 3) stage transfer in  assisted reproductive technologies . The Cochrane study showed a small improvement in live birth rate per couple for blastocyst transfers. This would mean that for a typical rate of 31% in clinics that use early cleavage stage cycles, the rate would increase to 32% to 41% live births if clinics used blastocyst transfer. [ 47 ]  Recent systematic review showed that along with selection of embryo, the techniques followed during transfer procedure may result in successful pregnancy outcome. The following interventions are supported by the literature for improving pregnancy rates:


•
Abdominal ultrasound guidance for embryo transfer


•
Removal of cervical mucus


•
Use of soft embryo transfer catheters


•
Placement of embryo transfer tip in the upper or middle (central) area of the uterine cavity, greater than 1 cm from the fundus, for embryo expulsion


•
Immediate ambulation once the embryo transfer procedure is completed [ 48 ]


Embryo transfer techniques allow top quality female  livestock  to have a greater influence on the genetic advancement of a herd or flock in much the same way that  artificial insemination  has allowed greater use of superior sires. [ 49 ]  ET also allows the continued use of animals such as competition mares to continue training and showing, while producing  foals . The general epidemiological aspects of embryo transfer indicates that the transfer of embryos provides the opportunity to introduce genetic material into populations of livestock while greatly reducing the risk for transmission of infectious diseases. Recent developments in the sexing of embryos before transfer and implanting has great potential in the dairy and other livestock industries. [ 50 ]


Embryo transfer is also used in  laboratory mice . For example, embryos of genetically modified strains that are difficult to breed or expensive to maintain may be stored frozen, and only thawed and implanted into a  pseudopregnant  dam when needed.


On February 19, 2020, the first pair of Cheetah cubs to be conceived through embryo transfer from a surrogate cheetah mother was born at Columbus Zoo in Ohio. [ 51 ]


The development of various methods of  cryopreservation  of  bovine   embryos [ 52 ] [ 53 ]  improved embryo transfer technique considerably efficient technology, no longer depending on the immediate readiness of suitable recipients. Pregnancy rates are just slightly less than those achieved with fresh embryos. [ 54 ]  Recently, the use of  cryoprotectants  such as  ethylene glycol  has permitted the direct transfer of bovine embryos. [ 55 ] [ 56 ]  The world's first live  crossbred  bovine calf produced under  tropical  conditions by Direct Transfer (DT) of embryo  frozen  in ethylene glycol freeze media was born on 23 June 1996. Dr. Binoy Sebastian Vettical of Kerala Livestock Development Board Ltd has produced the embryo stored frozen in Ethylene Glycol freeze media by  slow programmable freezing  (SPF) technique and transferred directly to recipient  cattle  immediately after thawing the frozen straw in water for the birth of this calf. In a study, in vivo produced crossbred bovine embryos stored frozen in ethylene glycol freeze media were transferred directly to recipients under tropical conditions and achieved a pregnancy rate of 50 percent. [ 57 ]  In a survey of the North American embryo transfer industry, embryo transfer success rates from direct transfer of embryos were as good as to those achieved with  glycerol . [ 58 ]  Moreover, in 2011, more than 95% of frozen-thawed embryos were transferred by Direct Transfer. [ 59 ]


Evans v. the United Kingdom  was a key case at the  European Court of Human Rights . The case outcome could have had a major impact on fertility law, not only within the  United Kingdom  but also the other  Council of Europe  countries.


Professor John Harris of the  University of Manchester  told the  BBC  in September 2002:


If the woman (Natallie Evans) succeeds in this case then the whole basis upon which the Human Fertilisation and Embryology Authority has operated thus far will be overturned. Until now, it has operated on the basis that there must be continuing consent between a man and a woman in every stage of the reproductive process. If she (Ms Evans) succeeds in this case, then she will have established that the man's role ends once the egg is fertilised. [ 1 ]


On 10 April 2007 Natallie Evans lost her final appeal at the Grand Chamber of the  European Court of Human Rights . [ 2 ]


In June 2000,  Natallie Evans  (born October 1971) and  Howard Johnston  (born November 1976) who had met while working at  Virgin Mobile  in  Wiltshire  became  engaged . In October 2001, Evans was diagnosed with  ovarian cancer , and was offered a cycle of  IVF  treatment before her cancer treatment because her fertility would be affected. On 12 November 2001 eleven of Evans' eggs were produced and fertilised using Johnston's sperm, resulting in six  embryos  which were frozen and placed in storage. On 26 November 2001 Evans underwent an operation to remove her  ovaries . She was told she would need to wait for two years before the implantation of the embryos in her  uterus . [ 3 ]


In May 2002, the couple split up. In the summer of 2002, Johnston, who lives in  Cheltenham , wrote to the  Bath, Somerset  clinic storing the embryos and asked that they be destroyed.


On 30 July 2002 the clinic informed Evans of Johnston's request under current United Kingdom IVF law, regulated by the  Human Fertilisation and Embryology Authority , which states that both parties must give their consent for IVF to continue – otherwise, the embryos must be destroyed.


Evans started her legal challenge immediately, which in agreement with her solicitor Muiris Lyons [ 4 ]  was a dual challenge on the existing UK law under the  Human Rights Act 1998 , with  Lorraine Hadley . Hadley from  Baswich ,  Staffordshire , underwent IVF treatment but later got  divorced  from her husband Wayne, and wanted to use two of her stored embryos to try to get  pregnant . [ 5 ]


Lawyers for the two women submitted to the  High Court of Justice  in September 2002 that, as treatment was already under way, the men should not have the right to stop it. The women used the analogy that if they got pregnant naturally and the embryos were in their bodies, then their partners would have no say at all. [ 5 ]  In September 2002,  Elizabeth Butler-Sloss, Baroness Butler-Sloss  the president of the High Court's Family Division, said that a full hearing would be scheduled for the New Year, later confirmed as 30 June 2003. [ 6 ]


On 1 October 2003 Mr Justice Wall said that in ruling against the women, although he had sympathy for the women's situation, he could not overrule the law as it stood. He said it was up to Parliament, rather than the High Court, to decide whether the law in this area needed to be changed. The High Court ruled the embryos of both women should be destroyed, but that would not happen until the conclusion of the appeals process. The women could appeal to the Court of Appeal or on to the European Courts. The main secondary outcome of the ruling was that the judge ruled that fertility clinics would now have to counsel couples having IVF to consider carefully what would happen to any embryos that were created if they split up. [ 7 ]


James Grigg, solicitor for Howard Johnston, said: "Mr Johnston firmly believes that this outcome could be the only one, given the circumstances. With the conclusion of his relationship with Ms Evans, he would not now elect to start a family with her. If a child had been born, Mr Johnston would have legal, financial, emotional and moral responsibility for it. Mr Johnston hopes that Ms Evans wish to start a family can be satisfied in some other way, perhaps using donor eggs." After offering to donate the embryos to infertile couples, [ 8 ]  on 24 October 2003 Mrs Hadley told the  BBC   Midlands Today  regional news program that she was giving up her fight in light of the unlikely scenario that her ex-husband would provide consent to her using the embryos, and the withdrawal of  legal aid . [ 9 ]


On 29 November 2004 three  Law Lords  dismissed Evans's application for leave to the  House of Lords  on the grounds that the petition "did not raise an arguable point of law of general public importance which ought to be considered by the House at this time, bearing in mind that the cause has already been the subject of judicial determination". [ 10 ]  Ms Evans still had the right to file a complaint to the European Court of Human Rights (ECHR) in  Strasbourg .


On 26 January 2005 the Bath clinic informed Evans that it was under a legal obligation to destroy the embryos, and intended to do so on 23 February 2005. Although now outside the statutory five year legal limit for the retention before use of embryos, on 27 February 2005 the ECHR, to whom Evans had applied, requested, under Rule 39 (interim measures) of the Rules of Court, that the United Kingdom Government take appropriate measures to prevent the embryos being destroyed by the clinic before the Court had been able to examine the case. The embryos were not destroyed.


On 7 March 2006 a panel of seven judges of the ECHR delivered a majority 5–2 ruling against Ms Evans, which read: "The Court, like the national courts, had great sympathy for the plight of the applicant who, if implantation did not take place, would be deprived of the ability to give birth to her own child." [ 11 ] [ 12 ]  However, the panel majority found that, even in such exceptional circumstances as Ms Evans', the right to a family life – enshrined in article eight of the European Convention of Human Rights – could not override Johnston's withdrawal of consent. The panel also ruled, this time unanimously, that the issue of when the right to life begins "comes within the margin of appreciation which the Court generally considers that States should enjoy in this sphere", and thus rejected the claim that embryo's right to life was being threatened. [ 13 ] [ 14 ]


Michael Wilks, of the  British Medical Association  ethics committee, said: "It's the right verdict, but a terrible situation." Dr Wilks called for a change to the five year limit for embryos to be stored after one partner withdraws consent should be extended so there was less of a "ticking clock". Ms Evans decided to appeal to the Grand Chamber of the European Court, but commented that she still wanted her ex-fiancé to change his mind to allow the embryos to be used. [ 13 ]


On 10 April 2007 the Grand Chamber of the ECHR ruled against Evans' appeal under three articles of the European Convention of Human Rights, which represented her last chance to save the embryos. The court ruled unanimously that there had been no breach of the right to life; but on the right to respect for private and family life and on the prohibition of discrimination the 17 judges ruled 13 to 4 (Judges Türmen, Tsatsa-Nikolovska, Spielmann and Ziemele dissented the judgment). [ 15 ]  Dr Allan Pacey, secretary of the British Fertility Society, said: "I think it was the only sensible decision which the Grand Chamber could come to. UK law is clear. It is a principle of shared responsibility." [ 2 ]


The timeline of the case is as follows: [ 16 ]


A  frozen zoo  is a storage facility in which genetic materials taken from animals (e.g.  DNA ,  sperm ,  eggs ,  embryos  and live tissue) are stored at very low temperatures (−196 °C) in tanks of liquid nitrogen. [ 1 ]  Material preserved in this way can be stored indefinitely [ 2 ]  and used for  artificial insemination ,  in vitro fertilization , embryo transfer, and  cloning . There are a few frozen zoos across the world that implement this technology for conservation efforts. Several different species have been introduced to this technology, including the Pyrenean ibex, Black-footed ferret, and potentially the white rhinoceros.


The first frozen zoo was established at the  San Diego Zoo  by pathologist  Kurt Benirschke  in 1972. [ 3 ] [ 4 ] [ 5 ]  At the time there was no technology available to make use of the collection, but Benirschke believed such technology would be developed in the future. [ 6 ]  The frozen zoo idea was later supported in  Gregory Benford 's 1992 paper proposing a Library of Life. [ 7 ]  Zoos such as the San Diego Zoo and research programs such as the Audubon Center for Research of Endangered Species [ 8 ] [ 2 ]  cryopreserve genetic material in order to protect the diversity of the gene pool of endangered species, or to provide for a prospective reintroduction of such extinct species as the  Tasmanian tiger [ 9 ]  and the  mammoth . [ 10 ]


Gathering material for a frozen zoo is rendered simple by the abundance of  sperm  in males. Sperm can be taken from an animal following death. The production of eggs, which in females is usually low, can be increased through  hormone treatment  to obtain 10–20  oocytes , dependent on the species. Some frozen zoos prefer to fertilize eggs and freeze the resulting embryo, as embryos are more resilient under the cryopreservation process. [ 11 ]  Some centers also collect skin cell samples of endangered animals or extinct species. The  Scripps Research Institute  has successfully made skin cells into cultures of special cells called induced pluripotent stem cells ( IPS cells ). It is theoretically possible to make sperm and egg cells from these IPS cells. [ 4 ]


Several animals whose cells were preserved in frozen zoos have been cloned to increase the  genetic diversity  of endangered species, as of 2021 [update] . One attempt to clone an extinct species was made in 2003; the newborn  Pyrenean ibex  died of a development disorder which may have been linked to the cloning, and there are not enough genetic samples in frozen zoos to re-create a breeding Pyrenean ibex population.


The Frozen Zoo at the San Diego Zoo's Institute for Conservation Research currently stores a collection of 8,400 samples from over 800 species and subspecies. [ 12 ]  It has acted as a forebear to similar projects at other zoos in the United States and Europe. [ 13 ] [ 14 ]  However, there are still less than a dozen frozen zoos worldwide. [ 2 ]


At the United Arab Emirates'  Breeding Centre for Endangered Arabian Wildlife (BCEAW) in Sharjah , the embryos stored include the extremely endangered  Gordon's wildcat  ( Felis silvestris gordoni ) and the  Arabian leopard  ( Panthera pardus nimr ) (of which there are only 50 in the wild). [ 11 ]


The Audubon Center for Research of Endangered Species, affiliated with the  University of New Orleans , is maintaining a frozen zoo. In 2000 the Center implanted a frozen-thawed embryo from the highly endangered African wildcat into the uterus of a domestic house cat, resulting in a healthy male wildcat. [ 15 ]


The  Frozen Ark  is a frozen zoo established in 2004 and jointly managed by the  Zoological Society of London , the  London Natural History Museum , and the  University of Nottingham . [ 16 ] [ 17 ]  This organization operates as a charity with many different departments including the DNA laboratory, consortium, taxon expert groups, and the database. [ 18 ]  In the DNA laboratory, samples are contained after collection from scientists, and different research projects are conducted there. The consortium acts as a bridge to bring together different, but important, groups from zoos, aquariums, museums, and universities. The taxon expert groups monitor the major phyla and lists like the IUCN Red List. The database is the essential piece as it holds all reports and records needed to perform all of the other functions for the charity. [ 18 ]  The hope for the future is for zoos and aquariums to be able to collect samples from their threatened and/or endangered species in house to help with conservation efforts. [ 18 ]  The collection and freezing of these samples allows for the distribution of gametes among populations. Samples can be collected from living hosts and from deceased hosts as well. [ 18 ] [ 19 ]


The  University of Georgia 's Regenerative Bioscience Center is building a frozen zoo. RBC Director Steven Stice and animal and dairy science assistant professor Franklin West created the facility with the thought of saving endangered cat species. The scientists have already extracted cells from a  Sumatran tiger , which could be used for artificial insemination. Artificial insemination provides a remedy for animals who, due to anatomical or physiological reasons, are unable to reproduce in the natural way. Reproduction of stored genetic material also allows for the fostering of genetic improvements, and the prevention of inbreeding. Modern technology allows for genetic manipulation in animals without keeping them in captivity. However, the success of their restoration into the wild would require the application of new science and a sufficient amount of previously collected material. [ 11 ]


Due to the very low temperatures required, varying levels of stress are put on the DNA samples.  Spermatozoa , in particular, are stressed by temperature shock, osmotic stress, and oxidative stress with the latter being the most detrimental. [ 20 ]  When temperature shock occurs, the membrane is damaged through freezing and thawing of the sperm. Osmotic stress occurs when ice crystals form inside the nucleus during the freezing process, causing differing osmotic pressures within the cell. Oxidative stress is the result of too many reactive oxygen species (ROS), which is highly reactive and damaging to all parts of the cell. [ 20 ] [ 21 ]  Although these stressors are present within the cell, there are solutions to each. By introducing cholesterol to the samples, temperature shock can be reduced. The use of antifreeze proteins provides one solution for osmotic stress. Oxidative stress is the most difficult to combat because of the highly reactive components of ROS, but some measures like adding certain proteins to limit freeze-thaw damage and increase the survival rate of the DNA. [ 20 ]


A  gaur  that died of natural causes had some skin cells frozen and added to the San Diego Frozen Zoo. Eight years later, DNA from these cells was inserted into a domestic-cow egg to create an embryo (trans-species cloning), which was then implanted in a  domestic cow  ( Bos taurus ). On 8 January 2001, the gaur, named Noah, was born in  Sioux Center, Iowa . Noah was initially healthy, but the next day, he came down with clostridial enteritis, and died of  dysentery  within 48 hours of birth. This is not uncommon in uncloned animals, and the researchers did not think it was due to the cloning. [ 22 ]


The  banteng  was the second endangered species to be successfully  cloned , and the first clone to survive beyond infancy. [ 23 ] [ 24 ]  Scientists at  Advanced Cell Technology  in  Worcester, Massachusetts , extracted DNA from skin cells of a dead male banteng, that were preserved in San Diego 's Frozen Zoo facility, and transferred it into  eggs  from domestic banteng cows, a process called  somatic cell nuclear transfer . Thirty  embryos  were created and implanted in domestic banteng cows. Two were carried to term and delivered by  Caesarian section . The first was born on 1 April 2003, and the second two days later. The second was  euthanized , apparently suffering from large offspring syndrome (an  overgrowth disorder ), but the first survived and lived for seven years at the San Diego Zoo, where it died in April 2010 after it broke a leg and was euthanized. [ 25 ] [ 26 ]


In 2020, the first cloned  Przewalski's horse  was born, the result of a collaboration between  San Diego Zoo Global ,  ViaGen Equine  and  Revive & Restore . [ 27 ]  The  cloning  was carried out by  somatic cell nuclear transfer  (SCNT), whereby a viable  embryo  is created by transplanting the  DNA -containing  nucleus  of a  somatic cell  into an immature egg cell ( oocyte ) that has had its own nucleus removed, producing offspring genetically identical to the somatic cell donor. [ 28 ]  Since the oocyte used was from a domestic horse, this was an example of interspecies SCNT. [ 29 ]


The somatic cell donor was a Przewalski's horse stallion named Kuporovic, born in the UK in 1975, and relocated three years later to the US, where he died in 1998. Due to concerns over the loss of  genetic variation  in the captive Przewalski's horse population, and in anticipation of the development of new cloning techniques, tissue from the stallion was  cryopreserved  at the San Diego Zoo's Frozen Zoo. Breeding of this individual in the 1980s had already substantially increased the genetic diversity of the captive population, after he was discovered to have more unique  alleles  than any other horse living at the time, including otherwise-lost genetic material from two of the original captive founders. [ 27 ]  To produce the clone, frozen skin  fibroblasts  were thawed, and grown in  cell culture . [ 30 ]  An oocyte was collected from a domestic horse, and its nucleus replaced by a nucleus collected from a cultured Przewalski's horse fibroblast. The resulting embryo was induced to begin division and was cultured until it reached the  blastocyst  stage, then implanted into a domestic horse  surrogate  mare, [ 30 ]  which carried the embryo to term and delivered a foal with the Przewalski's horse DNA of the long-deceased stallion.


The cloned horse was named Kurt, after Dr.  Kurt Benirschke , a  geneticist  who developed the idea of cryopreserving genetic material from species considered to be endangered. His ideas led to the creation of the Frozen Zoo as a genetic library. [ 31 ]  There is a breeding herd in the  San Diego Zoo Safari Park . [ 32 ]  Once the foal matured, he was relocated to the breeding herd at the  San Diego Zoo Safari Park , [ 32 ]  so as to pass Kuporovic's genes into the larger captive Przewalski's horse population and increase the genetic variation of the species. In 2023, a second horse, named Ollie, was cloned from the same cell line. [ 27 ]


To help mitigate  inbreeding depression  for two endangered species, the  Black-footed ferret (Mustela nigripes) ,  Revive & Restore  facilitates on-going efforts to clone individuals from historic cell lines stored at the San Diego Zoo Wildlife Alliance Frozen Zoo. The program seeks to restore genetic variation lost from the living gene pool.


On December 10, 2020, the world's first cloned  black-footed ferret  was born. This ferret, named  Elizabeth Ann , marked the first time a U.S. endangered species was successfully cloned. [ 33 ] [ 34 ]


The cells of two 1980s wild-caught  black-footed ferrets  that never bred in captivity were preserved in the San Diego Wildlife Alliance Frozen Zoo. One of them was cloned to increase genetic diversity in this species in December 2020. More clones of both are planned. They will initially be bred separately from the non-cloned population. [ 35 ]


The  Pyrenean ibex  went extinct in 2000. In 2003 frozen cells from the last one (a female killed by a falling branch) were used to clone 208 embryos, of which 7 successfully implanted in goats, and one made it to term. That one ibex died of respiratory failure just after birth; quite possibly as a result of the cloning process, its lungs had not developed properly. There may not be enough individuals' cells preserved to create a breeding population. [ 36 ] [ 37 ]  Despite the death of the ibex, DNA analysis revealed that the offspring was a legitimate clone from its last living descendent. [ 38 ]


Over the years, concerns over population declines of the  northern white rhinoceros  ( Ceratotherium simum cottoni ) have increased with the increasing value of their horns to poachers. Specifically, the population has declined nearly seventy percent from 2011 to 2019. [ 39 ]  Processes like SCNT can help aid in conservation efforts towards the revival of their population. Researchers are looking towards induced pluripotent stem cells (iPSC), as they hold limitless possibilities. [ 40 ]  With the lack of natural mating occurring within the species due to the limited number of them, this sub-species provides researchers the opportunity for iPSC intervention. Other methods, including artificial insemination with fresh semen (AI), have been used successfully in another sub-species, the Southern White Rhinoceros ( Ceratotherium simum simum ). [ 41 ]  Frozen-thawed semen has been tested and has seen some successes, helping solve issues with reproduction of the species as a whole. [ 41 ]


The  Genetics & IVF Institute  (GIVF) is an international provider of  infertility  and  genetics  services and products, and also engages in biomedical research in these fields. The Institute was founded in 1984 by Dr.  Joseph D. Schulman  and associates.  GIVF headquarters are in Fairfax, VA, US, and its facilities include locations in Pennsylvania, Minnesota, California, and Texas in the United States, as well as in China, Mexico, and several other countries.


Genetics & IVF Institute has been responsible for numerous key innovations to help couples have healthy babies and to improve the reproductive health of women. These include:


First U.S. medical center to introduce non-surgical ultrasound guided egg retrieval ( transvaginal oocyte retrieval ) for  IVF ; [ 1 ]
achieved America's first frozen  embryo  twin birth; [ citation needed ]
first American team to publicly report pregnancies using ICSI ( intracytoplasmic sperm injection ); [ 2 ]
first to offer clinical laboratory testing for certain mutations in the  BRCA1  and  BRCA2  genes which cause hereditary  breast cancer ; [ citation needed ]
developed the important technique of non-surgical sperm aspiration for improved treatment of  male infertility ; [ 3 ]
first in the world to initiate ovarian  cryopreservation  for storage of  oocytes  for women with cancer; [ 4 ]
discovered a method (non-disclosing preimplantation  genetic testing ) for the effective prevention of  Huntington disease ; [ 5 ]
developed MicroSort, an important flow-cytometric  human sperm  separation technique for preconceptual  gender selection . [ 6 ]


The services and research activities at Genetics & IVF have been the subject of articles in newspapers, such as the  New York Times . [ 7 ]


In 1998, the Institute opened a branch facility in  Shanghai , China.


The Institute's American activities involve a large donor egg program, a group of  sperm banks  and  egg banks , preimplantation genetics testing centers, and oocyte  cryopreservation  via  vitrification  in addition to more conventional IVF, infertility, and genetic services.


Norbert Gleicher  (born August 20, 1948) is an American  obstetrician - gynecologist  active in  obstetrical practice ,  in vitro  fertilization ,  reproductive endocrinology , and  reproductive immunology . [ 1 ]  He is a fellow of the American College of Obstetricians and Gynecologists (FACOG) and the American College of Surgeons (ACS) and currently serves as president, medical director and chief scientist of the  Center for Human Reproduction  (CHR) in New York City, a clinical fertility center that he founded in 1981. Simultaneously, he is President of the Foundation for Reproductive Medicine, a not-for-profit research foundation. [ 2 ]  Gleicher maintains additional academic appointments at  Rockefeller University , and  Medical University of Vienna . [ 3 ]


Gleicher was born in the Polish city of  Kraków . [ 1 ]  In his youth he moved to  Vienna , the capital of Austria, where he began his medical studies at the  Medical University of Vienna  in 1966. He completed his medical degree and rotating internship at the  Sackler Medical School  of  Tel Aviv University  in 1974. He later moved to  New York City  for a 6-month fellowship in immunology. There he completed his residency in obstetrics and gynecology in 1979 at the  Mount Sinai Medical Center . During his residency he also maintained a grant-funded research laboratory, mostly pursuing research in reproductive immunology. While still a  chief resident , Gleicher organized at Mount Sinai Medical Center an international congress on reproductive immunology, which led to the formation of the American Society of Reproductive Immunology (ASRI), with him being elected as the society's vice-president, and with the immunology conference becoming an annual event under the society's sponsorship. [ 4 ]  He was awarded the Dr. Solomon Silver Award in Clinical Medicine for his "ability to apply the advance in research to the practice of clinical medicine". [ 5 ]


In 1979, Gleicher was appointed assistant professor in the Department of Obstetrics and Gynecology at Mount Sinai, [ 6 ]  division director of the Division of Reproductive Immunology and director of undergraduate (medical student affairs) and graduate education (residency program) in the department. He served in all of these functions until 1981. [ 5 ]


Recruited to Chicago in 1981 as chairman of obstetrics and gynecology at Mount Sinai Hospital and professor of obstetrics and gynecology and immunology/microbiology at  Rush Medical College , [ 7 ]  he was put in charge of rebuilding an academic teaching department in the inner city. His department established the first  in vitro  fertilization (IVF) program in the city of Chicago and the Midwest. [ 8 ]  Gleicher traveled for training to René Frydman's Paris-based IVF program in preparation for the program's establishment. As the number of programs in the U.S. was still minuscule then,  The Chicago Tribune  dedicated a two-day series of articles to its opening. [ 9 ]  In 1982 was appointed as founding editor-in-chief of  The Journal of In Vitro Fertilization and Embryo Transfer  ( JIVF-ET ), holding this position for approximately 20 years. [ 3 ]  In 1991 Gleicher was awarded  Austrian Decoration for Science and Art  for his work.


After 10 years of building a department, Gleicher resigned his chairmanship at Mount Sinai to concentrate on management of a rapidly growing IVF center, the Center for Human Reproduction (CHR). [ 10 ] [ 11 ] [ 12 ]  By the end of 1999, Gleicher was in charge of IVF centers in Chicago and New York City, splitting his time each week between the two cities. In 2003, he sold the Chicago operations, and moved full-time back to New York City, where he continues to pursue both clinical care and research. [ 13 ] [ 14 ]


While maintaining his role in clinical patient care and research at the Center for Human Reproduction in New York, Gleicher has continued his academic work as a professor or adjunct professor at educational institutions such as  Yale University , [ 15 ]   New York University ,  Rockefeller University , [ 16 ]  and the  Medical University of Vienna . [ 3 ]


Gleicher is a member of the  American College of Obstetricians and Gynecologists , the  American Society for Reproductive Medicine , and the  European Society of Human Reproduction and Embryology , among other institutions related to reproductive health and general medicine. As a speaker, Gleicher has lectured at major events worldwide. [ 3 ]  In 2009, he was commissioned to deliver the  Patrick Steptoe Memorial Lecture  in honour of the prominent British obstetrician and gynecologist before the British Fertility Society. [ 17 ]


Since the 1970s, Gleicher has published hundreds of scientific articles, abstracts and book chapters on reproductive endocrinology and infertility. He was one of the founders of the  American Journal of Reproductive Immunology , where he also served as editor-in-chief. For 30 years, he was editor of the Journal of In Vitro Fertilization and Embryo Transfer, now known as the  Journal of Assisted Reproduction and Genetics . [ 5 ]  Other publications in which he served as editor or member of the editorial board include  The Mount Sinai Journal of Medicine ,  International Journal of Obstetrics and Gynecology ,  Asian Pacific Journal of Reproduction  and  PLOS One , among others. [ 3 ]


Gleicher, in collaboration with his CHR colleague David H. Barad, introduced androgen supplementation by way of  dehydroepiandrosterone  (DHEA), to improve  in vitro  fertilization (IVF) outcomes in women with diminished ovarian reserve in 2005. [ 22 ]  However, none of his studies used randomized, controlled trials; results have been mixed for researchers who did. [ 23 ]


Gleicher has been criticized for recommending his own brand of DHEA to his patients without always informing them that he owns the brand. His brand costs much more than competing brands. [ 23 ]


Gleicher is divorced and the father of two daughters. In 2019, Gleicher was the subject of a social profile in  The New York Times . He lives on the  Upper East Side  of  Manhattan . [ 1 ]


The  history  of  in vitro fertilisation  (IVF) goes back more than half a century. In 1959 the first birth in a nonhuman mammal resulting from IVF occurred, and in 1978 the world's first baby conceived by IVF was born. As medicine advanced, IVF was transformed from natural research to a stimulated clinical treatment. There have been many refinements in the IVF process, and today millions of births have occurred with the help of IVF all over the world.


Prior to the development of IVF in humans,  Walter Heape , a physician and professor at the  University of Cambridge , was doing research on reproduction in animals. In 1891, Heape reported the first successful embryo transfer in a mammal. [ 1 ] [ better source needed ] [ 2 ]


As early as 1934,  Gregory Goodwin Pincus  and Ernst Vincenz Enzmann tried to perform IVF in rabbits. Although the pregnancy was successful, it was later determined that the fertilisation occurred in vivo (in the body). [ 3 ]


Miriam Menkin  was the first to extract an intact fertilised egg. [ 4 ]  In 1948,  John Rock  and  Miriam Menkin  retrieved over 800  oocytes , of which 138 were exposed to  spermatozoa  in vitro. They published their findings in the  American Journal of Obstetrics and Gynecology . [ 1 ] [ 5 ]  There was a transient biochemical pregnancy reported by Australian Foxton School researchers in 1953. [ 6 ] [ failed verification ]


In 1958,  Anne McLaren  and John Biggers published a landmark paper in the journal Nature, outlining how they had successfully grown mouse embryos in vitro and transferred them into female mice. This showed it was possible to mix a sperm and an egg outside a women's reproductive system and create a healthy embryo.


In 1959,  Min Chueh Chang  at the  Worcester Foundation , proved fertilisation in vitro was capable of proceeding to a birth of a live rabbit. Chang's discovery was seminal, as it clearly demonstrated that oocytes fertilised in vitro were capable of developing if transferred into the uterus. [ 7 ]


The first pregnancy achieved through in vitro human fertilisation of a human oocyte was reported in  The Lancet  from the  Monash University  team of  Carl Wood , John Leeton and  Alan Trounson [ 8 ]  in 1973, although it lasted only a few days and would today be called a biochemical pregnancy.  Landrum Shettles  attempted to perform IVF in 1973, but his departmental chairman interdicted the procedure at the last moment. [ 9 ] [ 10 ]  There was also an  ectopic pregnancy  reported by  Patrick Steptoe  and  Robert Edwards  in 1976. [ citation needed ]


Steptoe and Edwards started collaborating in human IVF research in 1968. [ 3 ]  In 1977, Steptoe and Edwards successfully carried out a pioneering conception which resulted in the birth of the world's first baby to be conceived by IVF,  Louise Brown , on 25 July 1978 in  Oldham General Hospital , Greater Manchester, UK. [ 11 ] [ 12 ] [ 13 ]  Her younger sister, Natalie Brown, was the first person conceived with IVF to have children of her own, in 1999. [ 14 ] [ 15 ] [ 16 ]


In October 1978, it was reported that  Subash Mukhopadhyay , a physician from  Hazaribagh , India was performing experiments on his own with primitive instruments and a household refrigerator and this resulted in a test tube baby, later named as "Durga" (alias Kanupriya Agarwal) who was born on 3 October 1978. [ 17 ]  However, state authorities prevented him from presenting his work at scientific conferences [ 18 ]  and, in the absence of scientific evidence, his work is not recognised by the international scientific community. These days, however, Mukhopadhyay's contribution is acknowledged in works dealing with the subject. [ 19 ] [ better source needed ]


Steptoe and Edwards were responsible for the world's first confirmed boy conceived by IVF, Alastair Montgomery, born on 14 January 1979 in Glasgow. [ 20 ]  A team led by  Ian Johnston  and Alex Lopata were responsible for Australia's first baby conceived by IVF, Candice Reed, born on 23 June 1980 in  Melbourne . [ 21 ] [ verification needed ]


It was the subsequent use of stimulated cycles with clomiphene citrate and the use of human chorionic gonadotrophin (hCG) to control and time oocyte maturation, thus controlling the time of collection, that converted IVF from a research tool to a clinical treatment.


This was followed by a total of 14 pregnancies resulting in nine births in 1981 with the Monash University team.  Howard W. Jones  and  Georgeanna Seegar Jones [ 22 ]  at the  Eastern Virginia Medical School  in  Norfolk, Virginia , further improved stimulated cycles by incorporating the use of a follicle-stimulating hormone (uHMG). This then became known as controlled ovarian hyperstimulation (COH). Another step forward was the use of gonadotrophin-releasing hormone agonists (GnRHA), thus decreasing the need for monitoring by preventing premature ovulation, and more recently gonadotrophin-releasing hormone antagonists (GnRH Ant), which have a similar function. The additional use of the oral contraceptive pill has allowed the scheduling of IVF cycles, which has made the treatment far more convenient for both staff and patients.


The ability to freeze and subsequently thaw and transfer embryos has significantly improved the feasibility of IVF use. [ 23 ]  In 1983,  Alan Trounson  and Linda Mohr reported the first pregnancy which used  embryo cryopreservation  (frozen human embryos). However, this embryo was not carried to term. In December 1983, the first baby was born using this method which happened to be twins. [ 24 ]


The ability to freeze and preserve a woman's eggs to be used at a later date has also had impacts on IVF use. In 1986,  Dr. Christopher Chen  reported the first pregnancy which used  oocyte cryopreservation  (frozen eggs). The ability to freeze sperm has also been possible for generations. Both of these options have been beneficial in IVF use to allow people who must undergo radiation or menopause to still have children. [ 25 ]


Another important development in IVF was when  egg donation  became publicly available. The first pregnancy from egg donation occurred in 1983. In the United States, the first child born from egg donation occurred on February 3, 1984. This has allowed those who are unable to conceive a child due infertility or lack of eggs to become parents. [ 26 ]


Intracytoplasmic sperm injection  (ICSI) of single sperms was developed by  Gianpiero D. Palermo  et al. in Brussels ( UZ Brussel ), 1992. [ 27 ]  This has enabled men with minimal sperm production to achieve pregnancies. ICSI is sometimes used in conjunction with sperm recovery, using a testicular fine needle or open testicular biopsy. Using this method, some men with Klinefelter's syndrome, and so would be otherwise infertile, have been able to achieve pregnancy. [ 23 ] [ 28 ]  Thus, IVF has become the ultimate solution for most fertility problems, moving from tubal disease to male factor, idiopathic subfertility, endometriosis,  advanced maternal age , and anovulation not responding to ovulation induction.


Robert Edwards  was awarded the 2010  Nobel Prize in Physiology or Medicine  "for the development of  in vitro  fertilization". [ 29 ]   Steptoe and  Jean Purdy  were not eligible for consideration as the Nobel Prize is not awarded posthumously. [ 30 ]   Carl Wood  was dubbed "the father of IVF ( in vitro  fertilisation)" for having pioneered the use of frozen embryos. [ 31 ]


In the US, ART cycles started in 2006 resulted in 41,343 births (54,656 infants), which is slightly more than 1% of total US births. [ 32 ]


The Society for Assisted Reproductive Technology (SART) reported that in 2012 alone IVF resulted in about 61,740 babies born in the United States. The CDC estimates that IVF results in about 1 to 2 percent of births in the United States every year. [ 33 ]


In 2016, IVF resulted in an estimated 76,930 live-born infants in the United States. As of 2015 over one million babies have been born as a result of IVF in the U.S. and over 8 million worldwide. [ 34 ]


Horse cloning  is the process of obtaining a horse with  genes  identical to that of another horse, using an artificial  fertilization  technique. Interest in this technique began in the 1980s. The Haflinger foal  Prometea , the first living cloned horse, was obtained in 2003 in an Italian laboratory. Over the years, the technique has improved. It is mainly used on high-performance but castrated or infertile animals, for reproductive cloning. These horses are then used as breeding stock. Horse cloning is only mastered by a handful of laboratories worldwide, notably in France, Argentina, North America and China. The technique is limited by the fact that some differences remain between the original and its clone, due to the influence of  mitochondrial DNA .


Reproductive cloning of the  Pieraz  and  Quidam de Revel  horses began in 2005. The  International Federation for Equestrian Sports  (FEI by its acronym in French) decided to ban clones from competition in 2007, before authorizing them in 2012. A few clones are used in  equestrian sports , winning major titles such as the Argentine  polo  championship in 2013. Nevertheless, the number of cloned horses is growing every year. The practice is highly controversial, particularly for  bioethical reasons , since it involves a high failure rate on  embryos . It also raises questions about the management of horses'  genetic diversity , the future of the  horse breeding  profession, and the outbreak of new  genetic disorders  or fraud.


The horse is the seventh  species  to be cloned yet. [ 1 ]


Horse cloning has undergone a rapid qualitative and quantitative evolution. [ 2 ]  While Italian professor Cesare Galli believes that horse cloning has aroused less interest than that of other large mammals, [ 3 ]  other scientists believe that the high commercial value attained by some horses has created immediate interest, unlike in the case of less valuable agricultural animal species. [ 4 ]  Equine cloning owes much of its development to the Belgian stud farm of  Zangersheide , one of the pioneers of  artificial insemination  and  embryo transfer . According to Éric Palmer, a French biologist specializing in horse reproduction (who introduced  ultrasound  to mares and produced the first foal by  in vitro fertilization ), [ 5 ]  the way for the use of cloning was initiated in the 1980s by  veterinarian  surgeon Dr. Leo de Backer. He was in contact with some of the world's leading sports stables. According to Palmer, the ones that are interested are  Alwin Schockemöhle ,  Jan Tops , Thomas Fruhman,  John  and  Michael Whitaker ,  Willi Melliger , Jean-Claude Van Geensbergen and La Silla (in Mexico), among many others. [ 6 ]  The value of cloning high lineage horses was recognized as early as 1998, with the Westhusin study. [ 7 ]  Research to this end was publicly announced in 2001. [ 8 ]  That same year, with the support of  Genopole , Éric Palmer founded Cryozootech, a company dedicated to preserving the genes of horses with exceptional performance, with a view to future cloning. The horse is not the first large mammal to be cloned, as  Dolly  the sheep and other animals precede it, making it the seventh mammal to be cloned. [ 9 ]


The birth of three cloned mules in the United States on May 4, 2003, came just before that of the first horse. [ 10 ]  The first successful attempt to produce a viable clone was made by the Italian laboratory LTR-CIZ, which gave birth to  Prometea  on May 28, 2003, a  Haflinger  foal carried to term by her mother, whose genetic copy she is. [ 9 ]  Her birth was announced publicly on August 6, 2003. Born 36 kilogram after a natural delivery and a full-term pregnancy in Laboratory of Reproductive Technology,  Cremona ,  Italy , [ 11 ] [ full citation needed ]  At 2 months old, Prometea weighed 100 kg (220 lb) [ citation needed ]  The name "Prometea" is the  feminine  form of Prometeo (" Prometheus " in  Greek ). These scientists worked under the guidance of Professor Cesare Galli. [ 9 ]


Dr. Cesare Galli and others at the lab experimented with 841 reconstructed embryos; of the 14 viable embryos, four were implanted in surrogate mothers - only that of Prometea succeeded in being born. Prometea was born to her twin mother who her cloning cells originated from. [ 12 ]   Texas A&M University  was also undertaking a horse-cloning project when the Italian team first succeeded.


In 2002, LTR-CIZ merged with Cryozootech. In Italy, they produced the world's second cloned horse,  Pieraz-Cryozootech-Stallion . This is a purely commercial clone, aimed at obtaining a fertile genetic copy of a successful but castrated horse. According to  Bernard Debré , the birth of Pieraz-étalon heralded the commercial direction that equine cloning would later take. [ 13 ]  Prometea and  Pieraz  were obtained using the same method, that of Professor Galli. [ 10 ] [ 14 ]


Shortly afterwards, on 13 March 2005, Dr Katrin Hinrichs gave birth to Paris-Texas, a clone of  Quidam de Revel , in a laboratory at  Texas A&M University  in Texas (USA). The clone foal is also produced commercially for breeding purposes, at the request of  Quidam's  owner. The technique used is slightly different from that of the Italians. [ 15 ]  As a result, the number of clones produced has increased over the years. In 2009, the E.T. FRH clone became the first cloned  show jumping  horse authorized for breeding by a  studbook  (the  Zangersheide  studbook), while endurance champion  Pieraz's  clone entered its third breeding season. [ 16 ]  In Argentina, polo player  Adolfo Cambiaso  uses Crestview Genetics to clone his  polo ponies . At the end of 2010, a clone of his polo mare Cuartetera was sold at auction for a record  $ 800,000. [ 17 ] [ 18 ]  In May 2013, a non-clone foal was born for the first time from two parents cloned by  embryo transfer . [ 19 ] [ 20 ]  On 7 December 2013, a cloned  polo pony  won a major sporting competition for the first time. It was the Argentine polo championship. [ 21 ]  In 2018, equine cloning was widely used in Argentina's polo scene. [ 22 ]  The Argentine polo horse has become the most cloned animal in the world. [ 23 ]


Cloning research is often carried out in secret, due to poor public acceptance. [ 19 ]  Commercial cloning companies sometimes reveal the births of these horses, but the techniques employed remain mostly secret. [ 24 ]  According to the French national stud farm, the cloning technique used, known as "somatic" cloning, involves taking cells by  biopsy , usually from the breast of an adult animal.  Fibroblasts  are extracted and cultured in vitro until a sufficient number is obtained, then stored in  liquid nitrogen .  Oocytes  are harvested from a mare, either living or dead. The DNA is removed by  enucleation , then in vitro culture makes it suitable for receiving fibroblast DNA from the animal to be cloned. [ 25 ] [ 26 ]  Due to the high demand for mare oocytes, these are usually obtained from slaughterhouses. [ 3 ]


After a week or so of in vitro culture, the resulting  embryo  is implanted into the  uterus  of a carrier mare, using the  embryo transfer  technique. After eleven months'  gestation , the mare gives birth to the cloned foal. However, this type of gestation is much riskier than a conventional one. [ 25 ]


According to a Belgian researcher interviewed by  Le Vif , the failure rate is the main reason for opposition to cloning, for  bioethical reasons  due to the mortality of  embryos , fetuses and newborn  foals . [ 27 ]  This rate is high, but is gradually decreasing thanks to better-controlled techniques. Professor Galli obtained 15% viable embryos for his second clone,  Pieraz , compared with only 3% for  Prometea , [ 28 ]  the first horse, which required 328 attempts. [ 29 ]  The first  mule  trials, in 2003, involved 118 embryos, 13 of which produced a gestation, resulting in 3 live mules. [ 24 ]  To clone  Calvaro V  in 2006, Cryozootech used over 2,000  oocytes , which produced 22 embryos, only one of which was carried to term. [ 30 ]


Estimates of this rate vary from source to source. In 2012, according to a Belgian researcher, the average success rate for animal cloning was around 5%. [ 27 ]  Argentine researchers estimate that 6 or 7 embryos are needed out of 20 trials (in 2013). [ 31 ]  In 2010, according to a French source, around 2,500 mare oocytes had to be used to obtain a single viable foal. [ 26 ]  There are many abortions as well. Despite increased susceptibility to neonatal disease, a clone has the same life expectancy and robustness as a conventional horse. [ 32 ]  There is nothing to differentiate a clone from a conventionally bred horse. [ 4 ]


The cost of equine cloning varies between €200,000 [ 32 ]  and €300,000, [ 33 ]  depending on the source. In 2010, clones intended for sporting competitions represented just 22% of operations. [ 34 ]  Cloning is therefore mainly carried out in Europe for the purpose of breeding high-performance horses. A  gelding  can be copied to ensure its progeny. [ 35 ]  The same applies to a stallion that has become too old to reproduce, or to a mare whose number of foals is naturally limited. [ 25 ]  The use of cloning relies heavily on the belief that DNA is the most important factor in competition performance. [ 36 ]  Anne Ricard's study estimates that, in equestrian disciplines ( show jumping ,  dressage  and  endurance ) where geldings represent around 40% of competitors, the use of reproductive clones will enable a genetic improvement of 4% per generation. Once their fertility has been established, their semen is frozen as for any other stallion. [ 34 ] [ 37 ]


Effective cloning remains very marginal [ 34 ]  due to its cost. [ 31 ]  In the US and Argentina, requests for equine cloning come mainly from  polo  players (who allow their mares to play all the seasons) and  Arabian horse  breeders. [ 10 ]  Cloning can also be used to preserve rare breeds threatened with extinction, [ 38 ] [ 39 ]  but customers' motivations are essentially commercial. [ 34 ]  Nevertheless, the discovery of a perfectly preserved prehistoric foal in Siberia (in 2018) augurs well for cloning trials by Russian and Korean researchers, to resurrect extinct equine breeds or species. [ 40 ]


If the foal is the genetic copy of its donor, the question of the influence of the  mitochondria  that remain present in the recipient oocyte is still open. [ 41 ]  Mitochondria represent only 1 or 2% of the genome, but could influence the clone's sporting performance. [ 25 ]  They are more important in the case of a mare than a stallion, since the mare transmits her mitochondria during reproduction, unlike the stallion. [ 42 ]  Similarly, the cloned horse is not necessarily a perfect copy of the donor in terms of  phenotype  and character. The  horse markings  may vary, and the character, depending less on genetics than on the influence of the mother and upbringing, may also turn out to be very different. The technique also has its limits when it comes to breeding, as the pattern sought in horses evolves over time. There is therefore little point in cloning a horse clone. [ 32 ]  It also takes a long time to implement, and the number of specialized laboratories and companies is limited. The Kheiron company in Argentina, for example, estimates its waiting time at eighteen years, with demand far outstripping supply. [ 31 ]  The ban on clones in a large number of stud books and in certain competitions also limits interest. [ 26 ]


A few companies are known for their specialization in commercial equine cloning: ViaGen, Replica Farms, Crestview Genetics, Kheiron and Cryozootech. Competition between these laboratories is fierce. [ 31 ]  French company Cryozootech is a pioneer in the field, having produced the first commercial clone in 2005. [ 34 ]  It has made the production of famous horse clones its specialty. [ 43 ]  ViaGen was originally based in  Texas  in the United States, but the laboratory moved to Canada after the last American  slaughterhouses  closed in 2007, to source mare  oocytes . [ 44 ]  Kheiron was set up in Argentina in 2009 with a team of eight people. Equine cloning has developed strongly in this country, thanks in particular to demand from  polo  players, the profusion of mare oocytes available for research (the country exports a lot of  horse meat , and has numerous slaughterhouses supplying oocytes) and the easy breeding conditions in the  Pampas . [ 31 ]  In 2012, Argentina was estimated to be the country producing the most horse clones in the world. [ 45 ]  In  Texas , more than 900 clones were born between the creation of the first laboratory and 2014. [ 31 ]  In 2019, equine cloning companies are expected to open in China. [ 46 ]


Canada since 2007 [ 44 ]


Argentina


Canada


The use of clones for the genetic improvement of horses' sporting performance is recognized, including by veterinarians, although skepticism remains high among some professionals. [ 2 ]  According to Éric Palmer, acceptance of horse cloning is growing, and attitudes are changing, in the same way as the gradual acceptance of  in vitro fertilization  and  artificial insemination  techniques in horses. [ 32 ]  Fears of the birth of malformed or monstrous animals diminished when clone owners realized that their animals were in good health. [ 52 ]


In 2007, the  International Federation for Equestrian Sports  ruled that cloned horses should be banned from the official competitions it organizes, believing that opening up participation to clones would be unjust and unfair to the competition. It revised its opinion in July 2012. Horse clones are now allowed in all FEI competitions. [ 53 ]  This reversal is seen as an important sign of recognition of the usefulness of clones in sport horse breeding. [ 37 ]


In the United States, the National Cutting Horse Association and the National Barrel Horse Association allow clones in  cutting  and  barrel racing  competitions. [ 34 ]  The  American Quarter Horse Association  was taken to court by owners and riders of cloned horses in 2012, for refusing to allow these horses to take part in official breed competitions. [ 21 ] [ 54 ]  The initial ruling ordered the association to amend its bylaws to allow clones to compete. [ 55 ]


Sporting training of the Levisto Alpha Z clone makes it possible, even probable, for a clone horse to win an Olympic title in the future. [ 56 ]


Clones are generally not entered in the stud books of their respective breeds. The American  Jockey Club  refuses to allow cloned horses to race. In France, clones are also banned from trotting and galloping races. [ 21 ]  Several European  sport horse  and  warmblood  studbooks accept clones:  Zangersheide ; Anglo-European (AES);  Irish Sport Horse  (ISH);  Dutch Warmblood  (KWPN);  Belgian Warmblood  (BWP); and  Holsteiner . [ 25 ] [ 57 ] [ 58 ]  France's  national stud farms  advise against banning clones from the various studbooks, arguing that this will ultimately drive the best gene pool abroad. [ 34 ]


The existence of clones is not always made public, due to the poor reception they receive. [ 10 ]  Although in Belgium, Isabelle Donnay believes that commercial cloning of horses has not been very successful, [ 27 ]  on a global scale, their numbers have clearly increased over time. Equidia Life's 2013 survey describes the practice as "booming". [ 19 ]  In winter 2010, 56 clones were counted worldwide, produced by laboratories in Europe, the United States and South America. Americans clone more mares than Europeans. [ 34 ]  Between 2006 and 2011, at least 20  American Quarter Horses  were cloned. [ 59 ]  In 2014, there were an estimated 900 clones in the state of  Texas . [ 10 ]  In Europe, the Belgian  Zangersheide  stable regularly uses this technique, with four horses cloned between 2006 and 2013. [ 33 ]  The stallion Salute, one of Smart Little Lena's clones, was exported to Australia in 2010 for breeding. [ 44 ]


Dr. Éric Palmer, Cryozootech, France


Dr.  Éric Palmer, Cryozootech, France


Poetin II Z ( Zangersheide )


Murka's Gem


Gem Twist Alpha Z ( Zangersheide )


July 2012


2008 (success)


Ratina Z Beta ( Zangersheide )


Ratina Z Gamma ( Zangersheide )


South America's first cloned horse


Jazz Clone 2


Cruising Encore


As Cold As Ice Beta


Cocaine Beta Z


Chilli Morning III (Trey)


Chilli Morning IV (Quattro)


Nintendo 64


According to various surveys, including one carried out by Cheval Savoir in 2009, horse cloning is generally poorly accepted by riders and horse professionals. They believe it introduces unfair competition to "normal" horse breeders, while constituting a highly lucrative and  ethically  unacceptable activity. For French scientist Éric Palmer, the technique is  demonized  due to misunderstandings. [ 32 ]  The  American Quarter Horse Association  has stated that "[...] clones have no parents, cloning is not breeding. It's just photocopies of the same horse", pointing to its low success rate and the risk of as yet unknown  genetic disorder  developing. [ 88 ]  The  Jockey Club  was also strongly opposed. [ 37 ]  Dr. Thomas Reed, who owns the private stud Morningside in Ireland, is also publicly opposed to cloning after the accidental death of his stallion  Hickstead  in competition at the end of 2011. [ 51 ]


In 2015, the  European Union  voted to ban the cloning of farm animals (cattle, pigs, sheep, goats, and horses), and the sale of cloned livestock, their offspring, and products derived from them, such as meat and milk. The ban excluded cloning for research, and for the conservation of rare breeds and endangered species. [ 89 ] [ 90 ]  However, no law was passed after the vote. As of 2024, horse cloning continues to be legal in the EU, with the  Zangersheide  registry in  Belgium  offering three cloned stallions for breeding. [ 91 ]


Horse cloning, like that of other animal species, raises bioethical issues, since it involves a high mortality rate of  embryos ,  fetuses  and young foals. The Swiss National Stud's ethics study reports "massive loss during gestation", with less than 1% of oocytes obtained resulting in a live foal. Furthermore, foals born from cloning suffer from frequent health problems. An American study looked at 14 clones born between 2004 and 2008. [ 92 ]  Six (43%) were normal, while the other eight suffered from neonatal disorders, umbilical problems and limb deformities. [ 92 ]  There are a large number of stillborn foals, deaths in the first few days after birth, immune deficiencies, and muscle and bone deformities. Problems at foaling are common for both carrier mare and foal, with  cesarean section  being a common option. If foals survive their postnatal period, they do not appear to be more susceptible to disease thereafter. The question of their  longevity  remains unknown, as the first clones are still too young to draw any statistics. [ 93 ]


In the UK, researcher William (Twink) Allen was refused permission to continue his cloning trials in 2004 for these ethical reasons, as the cloned animals could present malformations, anomalies and diseases, according to the British authorities. [ 94 ]  Dr. Natasha Lane, of the  Society for the Prevention of Cruelty to Animals  (SPCA by its acronym in French), said it was not acceptable to clone animals by sacrificing embryos "just to get a gold medal". [ 95 ]  Allen spoke out against the decision, saying that the British government had "caved in to the  animal protection  lobby". [ 28 ]


Although horses are not threatened with extinction or other major problem now, cloning may create less genetic diversity among horses by using these horses to breed.  This increases the life time of one breeding set of genetics resulting in less variability in a population. In conservation biology, there are concerns related to the lack of genetic diversity that allows for continuation of the species through genetic variation. [ 96 ]


On 8 June 2005, a number of French farmers belonging to the Confédération Paysanne demonstrated in front of the  Genopole  d' Évry , Cryozootech's headquarters, to denounce the "seizure of living matter" and a future loss of  genetic diversity , arguing that the development of cloning will eventually lead to the disappearance of the  breeding  profession. [ 10 ]  A number of specialists warn against the widespread use of cloning, believing that it will seriously damage the equine breeding industry, particularly in equestrian sport, by reducing demand for naturally-born foals. Cloning would also drastically reduce genetic diversity, as the same genes "could be reproduced over and over again". [ 97 ] [ 98 ]


One fear that has arisen with cloning is that of new forms of fraud. In studbooks that refuse to accept clones, particularly the  Thoroughbred  studbook, it would be possible to pass off a horse cloned from a champion as another animal by falsifying its identification documents. [ 97 ]


Adriana Iliescu  (born 31 May 1938) is a Romanian retired university teacher, philologist and author of children's novels. She received international media attention in 2005, when she gave birth to daughter Eliza at age 66, making her the oldest birth mother in the world until this record was broken in 2006 by  María del Carmen Bousada de Lara . Eliza, who was delivered at Giuleşti Maternity Hospital on 16 January 2005 in  Bucharest , is Iliescu's biological (but not genetic) child, since she gestated in Iliescu's womb but was conceived by an  ovum  and  sperm  donated anonymously. [ 1 ] [ 2 ] [ 3 ]


Iliescu was first given  hormone  treatment to reverse  menopause  in 1995 and  in vitro fertilisation  (three  zygotes  with  sperm  and  ovum  from two anonymous donors) in 2004, becoming pregnant with  triplets .  After ten weeks one of the three  fetuses  failed to progress and died.  The remaining two fetuses, both girls, weighed just 1.45 kilograms (3.19 pounds) and 0.69 kilograms (1.54 pounds) after 33 weeks of  pregnancy , but after complications the smaller of the two died in the  womb .  Though doctors were expecting to perform a  caesarian section  soon after the 34th week, the death of one of the twins led to the decision to operate earlier than planned.  The surviving baby was expected to remain in hospital for six weeks. [ 4 ]


Romanian laws governing the process are currently under review and, to bring them in line with typical European legislation, may prevent any form of such treatment after the age of 50.


The story became international  headline  news, causing debate as to whether fertility treatment is ethical after a certain age.  The release of details about the pregnancy and birth was criticised when different information was reported by different news companies.  One primary source was a  Realitatea TV  interview with Iliescu conducted a month before the birth.  The age of Iliescu was reported as 67 by some sources and the exact details of the second, and in some reports third, fetus differed greatly.  For example, a  CNN  news website article was updated after a day, changing her age from 67 to 66. [ 5 ] [ 3 ]  Gheorghe Borcean, head of the Romanian medical profession's ethics committee, commented: "A case of such prominence should require academic debates and not just one single television report." [ 2 ]


Television, newspaper polls and discussions around the world asked the public what their opinion of the ethical decision was. [ 6 ]


In vitro maturation  ( IVM ) is the technique of letting the contents of  ovarian follicles  and the oocytes inside mature  in vitro . It can be offered to women with infertility problems, combined with In Vitro Fertilization (IVF), offering women pregnancy without ovarian stimulation.


In 1935, Pincus & Enzmann did the first experiment on immature rabbit oocyte, showing  in vitro  spontaneous maturation and fertilization. [ 1 ]  They showed maturation occurs in isolation from normal follicular environment. [ 1 ]  In 1965 Edwards then continued IVM studies in mouse, sheep, cow, pig, rhesus monkey and human. [ 2 ] [ 3 ]  By 1991, the first human pregnancy was recorded using IVM followed by IVF, [ 4 ]  and in 1994 the first birth using IVM oocytes from  polycystic ovarian syndrome  patients was recorded highlighting that PCOS patient's oocytes are capable of maturation. [ 5 ]


Oogenesis  takes place during fetal life, in which primordial germ cells undergo  mitosis  until a few weeks prior to birth, forming  oogonia . These then begin meiosis to form the oocyte within the primordial follicle. [ 6 ]  This follicle consists of the oocyte surrounded by flattened pregranulosa cells. Babies are born with 1-2 million primordial follicles, and by puberty have around 300,000. [ 6 ]  Of these primordial follicles, only around 400 mature oocytes are released and could be potentially fertilised, with the rest undergoing atresia. [ 7 ]


'Maturation' of an oocyte is the process by which an 'oocyte attains the competence to be fertilised and undergo embryogenesis'. [ 8 ]


Folliculogenesis  is the mechanism by which the ovarian follicles mature. This can take many months in vivo and involves primordial follicle growth and differentiation. [ 8 ]


Primordial follicles containing the primary oocyte, arrested at prophase of meiosis I, [ 8 ]  develop into primary follicle containing cuboidal granulosa cells. A secondary follicle is formed with a few granulosa cell layers, as well as a theca layer. Finally before ovulation, a tertiary follicle is formed containing a follicular-fluid filled antrum. [ 6 ]  Of these small antral follicles, 1 will become dominant and ovulate (in monoovulatory species). During ovulation, the primary oocyte will resume meiosis in response to signals, arresting in metaphase meiosis II, ready for fertilization. [ 3 ]  The dominant follicle contains the mature oocyte. Follicular development is directly under gonadotropins control, LH and FSH. These use cAMP as an intracellular second messenger, with growth factors and cytokines also influencing their development in vivo. [ 7 ]


Through  in vitro  maturation, folliculogenesis and latter parts of oogenesis are being mimicked outside of the ovaries- trying to recreate the conditions for these processes.


If a follicle has reached the early tertiary or antral stage, IVM can be carried out. [ 9 ]


Firstly, the oocytes need to be obtained from the subject. The timing of this is dependent on the stage of the cycle the subject is in, which is usually monitored using  ultrasonography . [ 10 ]  If without the use of priming, oocytes are obtained when the largest follicles are around 10mm in size. [ 9 ]


In humans, this can be done with an aspiration needle, using ultrasound to allow accuracy. Depending on whether you are aspirating mature or immature follicles, the protocol differs slightly. In both procedures the aspiration pressure is reduced, but to varying degrees. Additionally, it is more important that the aspirate is filtered when retrieving immature follicles, as the follicles are much smaller and harder to see in the fluid extracted. [ 10 ]


Priming is the process by which the oocytes are primed with  follicle-stimulating hormone  (FSH) or  human chorionic gonadotrophin  (hCG) before retrieval. hCG is important in women with  polycystic ovarian syndrome  (PCOS). This results in an expanding or dispersed pattern of the  cumulus oophorus  around the egg cell, facilitating its identification within follicular fluid. This leads to improved maturation and quality of the oocytes. [ 7 ]  However, the evidence of a clinical effect of hCG priming is still lacking. [ 11 ]  When IVM was initially introduced, successful pregnancies were low, leading to the use of ovary priming. [ 10 ]


This technique is also used in sheep, [ 12 ]  pigs [ 13 ]  and other animals. See  In animals .


Oocytes are classified depending on their condition, such as number of cumulus cells. The best oocytes are chosen to be matured in the hope of then being implanted using  in vitro  fertilisation techniques. [ 12 ]


The oocytes are then cultured in media containing nutrients important for oocyte survival, such as gonadotrophins, growth factors and steroids. [ 10 ]  These vary between clinics and research laboratories. McLaughlin  et al.  biopsied human ovarian tissue and achieved a 10% rate of maturation from unilaminar follicles into  metaphase II  by a multi-step culture system: [ 14 ]


Once the oocytes have sufficiently matured, they can then be fertilised  in vitro , known as  in vitro fertilisation  (IVF). Techniques such as  intracytoplasmic sperm injection  (ICSI) can also be utilised to improve the chances of fertilisation being successful, which should be performed at least one hour (and optimally two to four hours) after the first  polar body  extrusion. [ 15 ]  Out of  in vitro  matured oocytes, those fertilised with ICSI have a success rates of 60-80%, compared to IVF with success rates of 25-40%. [ 16 ]


A few live births have already been made by taking small early tertiary follicles, letting them mature  in vitro  and subsequently  fertilizing  them.
However, for follicles that haven't reached the early tertiary stage, IVM is still under development. There are a lot of cellular changes in the  oocyte  and the rest of the cells in the follicle, which makes it very susceptible. Nevertheless, it is possible to let a primordial follicle mature to a secondary follicle outside the body by growing it in a slice of ovarian tissue. The subsequent maturity from secondary to early tertiary stage can then be supported in test-tubes. [ 16 ]  It has been suggested that photoirradiation of granulosa cells and oocytes may facilitate IVM. [ 17 ]


In vitro  maturation is an  assistive reproductive technique  ( ART ) typically used in patients with fertility issues including  polycystic ovary syndrome  (PCOS), high  antral follicle  counts and ovarian hyper-responsiveness. [ 18 ] [ 19 ]  However, more recently IVM has also become widely utilised in areas such as  fertility preservation  in cancer patient who have undergone treatment involving gonadotoxic therapies. [ 18 ]  There have been over 1000 live births recorded from mothers using IVM. [ 19 ]


PCOS is a common disorder involving dysfunction of the  endocrine system  associated with female reproduction. PCOS involves discrepancies in the  Hyphophyseal-pituitary-gonadal endocrine axis  which can result in hormonal dysfunction, excess  androgens  (e.g. testosterone) and frequent  anovulatory  menstrual cycles. [ 20 ]  Therefore, it is common for women suffering from PCOS to require assistance in order to conceive. [ 20 ] [ 21 ] [ 22 ]  In these patients IVM can be used to mature oocytes and aid conception. [ 20 ] [ 21 ]  Few studies shows that substituting IVM in PCOS patients eliminates the risk of OHSS and lowers the cost of treatment. The same group conducted a retrospective analysis study to compare the treatment outcome of IVM with IVF in patients with PCOS.They have further concluded there was significant increase pregnancy rates, implantation rates and number of embryos transferred in IVM group. [ 23 ]


The use of  in vitro  maturation in assisted reproduction has advantages over standard ART procedures. In typical  IVF  practice,  controlled ovarian hyperstimulation  is performed, which is where  supraphysiological  levels of  gonadotropins  are administered to the patient in order to hyperstimulate the antral follicles and hence induce oocyte maturation to  metaphase II  at a rate that is above normal physiological capabilities. [ 19 ]  This practice can be disadvantageous in several ways: It is very costly, can become complicated and may also predispose to several undesirable side effects, such as  ovarian hyperstimulation syndrome (OHSS) . [ 19 ] [ 21 ]  Ovarian hyperstimulation can cause severe OHSS in up to 2% of cases. OHSS can have serious consequences, including respiratory problems, renal impairment and even stroke. [ 19 ]  Patients with PCOS and younger women are at an increased risk of OHSS. [ 21 ]  In these women, it may be even more beneficial to employ IVM rather than conventional IVF treatment. [ 19 ] [ 21 ]


In IVM, immature oocytes are removed from the antral follicle of a woman and then are matured  in vitro  in a culture rich in gonadotrophins. [ 19 ]  This hence negates (or significantly reduces) the need for gonadotrophin stimulation. [ 21 ]


IVM is not an entirely perfected technique. Pregnancy rates are lower in IVM than in standard IVF. There is also research required into whether or not babies born to mothers who have undergone IVM have any health concerns (e.g. developmental issues) later in life. [ 19 ]


Women with a personal or family history of an o estrogen associated thrombus , or of severe cardiovascular disease, may also benefit from IVM. This is because conventional IVF, with its hyperstimulation of the ovaries, has the potential to stimulate mass synthesis of oestrogen via the stimulation of  granulosa cell  oestrogen production. [ 19 ]


Ovarian tissue cryopreservation  can be used as a method of  fertility preservation , such as before undergoing  chemotherapy  that can cause  female infertility , or as a future resource in case the oocytes will stop functioning by  advanced maternal age . Thus, ovarian tissue cryopreservation is an alternative to  oocyte cryopreservation  which requires a preceding  controlled ovarian hyperstimulation .  In vitro  maturation allows oocytes from the ovarian tissue to be used directly for  in vitro fertilization , as an alternative to surgical re-insertion of the tissue into the body. [ 14 ]


IVM may also be an important consideration for female patients diagnosed with  empty follicle syndrome  ( EFS ). In EFS, no oocytes are retrieved from mature ovarian follicles despite the application of supraphysiological levels of gonadotrophins. A woman can be diagnosed with EFS after she has undergone multiple rounds of IVF with total (or near total) failure in each round. [ 21 ]


Rescue IVM is a variant of classical  in vitro  maturation that involves attempting to mature immature oocytes that have been removed from a patient secondary to ovarian hyperstimulation in standard IVF practice.  Therefore, allowing for more oocytes to mature to the developmental stage where they can be developmentally viable. However, rescue IVM has been considered  a controversial field: If oocytes have not matured sufficiently in vivo – despite exposure to significant levels of gonadotrophins – it may be indicative of dysmaturity and of a limited potential developmentally. [ 19 ]


IVM has also been used in domestic animals including mice, [ 24 ]  cats, [ 25 ] [ 26 ]  dogs, [ 27 ] [ 28 ]  swine, [ 29 ]  sheep, [ 30 ]  horse [ 31 ]  and cattle [ 32 ] [ 33 ]  as well as wild species such as buffalo, [ 34 ]  bison, [ 35 ]  fish, [ 36 ]  lions, [ 37 ]  tigers [ 37 ]  and leopards. [ 37 ]  The ability to recover animals' oocytes initially destined for  ovarian follicle atresia , can be utilized by researchers, conservationists and the agriculture industry for academic purposes or for improving breeding systems.


In research, IVM can be carried out on animals so as to understand the developmental capacities of oocytes under certain conditions, or to understand the specific reproductive biology during that developmental period. IVM in other species is also carried out as some animals are used as models to study human-related reproductive biology. [ 38 ]  This research is often carried out with the aim of improving success rates of  in vitro  systems and/or aim to improve fertility in vivo.


Animal IVM models can be used to study how different exposures to harmful and toxic substances affect the maturing oocytes and their ability to become fertilized and develop into an embryo. [ 39 ]


It can also be used for subsequent biotechnology applications such as for the creation of  transgenic animals  using innovative gene-editing techniques such as  CRISPR/Cas9 ,  TALENs  and  ZFNs  for biomedical research. An example includes genetically engineered pigs with  CD163  and  CD1D  genes  knocked out . [ 40 ]  One of the ways these pigs were created was by injecting the CRISPR/Cas9 system into fertilised oocytes that were matured  in vitro .


In agriculture, IVM is usually carried out prior to IVF or artificial insemination as a means of conserving desirable traits of particular animals within herds and counteracting lower production as a result of seasonal breeding. In livestock species such cattle, transvaginal oocyte recovery from the ovaries of live female animals can be repeatedly carried out prior to the  in vitro  production of embryos. [ 41 ]


In non-domesticated animals, IVM can also be used for the conservation of endangered species whilst maintaining genetic diversity. [ 42 ]  However, due to limited resources and the species-specific nature of assisted reproductive technologies, the application of techniques such as IVM is still rare for non-domesticated animals. [ 42 ]


In an experiment by Segers I et al. (2015), the overall maturation rate after IVM of oocytes recovered from ovariectomy specimens in laboratory was 36%. The maturation rate correlated with the age of patient and duration of IVM. With the 8 couples with embryo  cryopreservation , there was a 65% fertilisation rate. At least one good quality day 3 embryo was cryopreserved in 7/8 couples. This experiment shows that IVM of oocytes obtained ex vivo during the processing of ovarian cortex prior to cryopreservation is a promising solution for patients at risk for fertility loss. [ 43 ]


The success of embryo production  in vitro  depends upon the use of an efficient oocyte retrieval technique and the best results have been obtained by laparoscopic aspiration. [ 44 ]


The obstetric and perinatal outcomes of births from IVM cycles are similar to those with  ICSI  treatments. [ 45 ]  However, IVM involves the use of invasive techniques, this may harm the mother. Furthermore, embryological outcome of IVM is not established. [ 46 ]  A more comprehensive appraisal of health status of IVM children will demand larger prospective studies. [ 45 ]  The potential of cryopreserved IVM oocytes from cancer patients remain unknown. The optimal number of IVM oocytes frozen in candidates for  fertility preservation  (FP) is unknown. FP oocytes  of infertile PCOS women have decreased competence compared to oocytes recovered after ovarian stimulation. The FP strategy of cryopreservation of oocytes after IVM should only be considered should ovarian stimulation is unfeasible. [ 47 ]


In norma-ovulatory women, the success rate of IVM is lower than conventional ovarian stimulation regimens with poorer implantation and pregnancy rates. IVM is suboptimal and influenced by several factors. However, IVM is a milder approach to assisted reproduction treatment and an alternative procedure for specific conditions. Accurate patient selection can improve IVM clinical outcome. [ 45 ]


IVM of oocytes cryopreserved may assist urgent fertility preservation in cancer patients. However, there is insufficient data regarding this outcome. Improving the culture conditions may increase the maturation rates and the potential of IVM oocytes. [ 48 ]


Besides that, in mouse oocytes, I-Carnitine (LC) supplementation during vitrification of  germinal vesicle  (GV) and their subsequent IVM improved nuclear maturation as well as meiotic spindle assembly and mitochondrial distribution in MII oocytes. However, no data to date has proven this benefit in fetal development and birth of healthy offspring after embryo transfer to surrogate females. However, this protocol could potentially improve the quality of vitrified human oocytes and embryos during IVM. [ 49 ]  In a research by Wang X et al. (2014), gonadotropins affect oocyte maturation, fertilisation and developmental competence in vitro. The responsiveness of bovine oocytes to gonadotropins in vitro depends on the relative concentrations (FSH/LH) for optimal oocyte development  developmental competence. Optimal FSH/LH concentrations could improve therapeutic clinical stimulation protocols and IVF success rates. [ 50 ]


The  Israeli IVF mix-up case  was a series of legal, ethical, and medical events following the discovery of a genetic error during an  in-vitro fertilization  (IVF) procedure at  Assuta Medical Center  in  Rishon LeZion , Israel. The incident, which began in September 2022, involved a pregnant woman who was informed that the fetus she was carrying was not genetically related to her or her partner. This discovery led to a highly publicized and contentious legal battle that spanned over two years, raising important questions about genetic parenthood, emotional bonds, and the best interests of children in similar situations.


In September 2022, a woman at Assuta Medical Center, while in her 30th week of pregnancy, was informed through  genetic testing  that the fetus she was carrying was not biologically related to her or her husband. Following this discovery, the  Israeli Ministry of Health  intervened and immediately began an investigation. A committee, led by Professor Ami Fishman, was formed to identify the biological parents of the fetus. The hospital was ordered to cease taking on new IVF patients to prevent similar occurrences until the investigation was complete. [ 1 ]


The baby was born on October 26, 2022, and the woman who gave birth to her chose to keep and raise her. The parents were officially registered as the baby's legal guardians in the  Israeli Ministry of Interior . However, despite their wishes, the medical center continued to search for the biological parents, resulting in a complex legal dispute.


On January 29, 2023, the Central District Court of Israel rejected an appeal by the non-biological parents and ordered genetic testing on several other couples and a single mother who claimed the child might be theirs. The  Supreme Court of Israel  decided on March 8, 2023, to halt further genetic testing on additional couples. Nevertheless, by August 2023, the Central District Court allowed a genetic test for another couple, escalating the legal complexities. The non-biological parents appealed to the High Court in an effort to prevent further testing.


On March 7, 2024, it was publicly revealed that six months prior, the biological parents of the child had been identified. This development led to continued disputes regarding custody and the legal status of the child. [ 2 ]


On November 24, 2024, the Family Court in Rishon LeZion made a final ruling that the two-year-old child, who had been raised by the non-biological parents since birth, should be transferred to her biological parents. [ 3 ]


Georgeanna Seegar Jones  (July 6, 1912 – March 26, 2005) was an American  reproductive endocrinologist  who with her husband,  Howard W. Jones , pioneered  in vitro fertilization  in the United States. [ 1 ]


She was born July 6, 1912, in  Baltimore, Maryland , to J. King Seegar. Her father was a obstetrician, one of the many things that led to Seegar Jones's interest in medicine from a young age. [ 2 ]  She was raised along with two siblings. She received her bachelor's degree in 1932 from Goucher College and continued to pursue her medical career at  Johns Hopkins University School of Medicine . [ 2 ]  Four years later, in 1936, she got her official medical degree (MD). [ 2 ]  She completed her training as a house gynecology officer and an acting member of the National Cancer Institute. [ 2 ]  While attending medical school she met her husband, Howard W. Jones, Jr., whom she married in 1940. [ 2 ]  Reproductive endocrinology was not yet a subspecialty; in fact, her and her husband's accomplishments have contributed to the successful reproductive endocrinology programs that are offered at Johns Hopkins. [ 2 ]


As a resident at Johns Hopkins, she discovered that the common pregnancy hormone, human chorionic gonadotropin,  hCG  was produced by the  placenta , not the  pituitary gland  as originally thought. [ 3 ] [ 4 ]   Thus leading to the development of the hCG pregnancy test that are currently on the market. By the late 1940s, Jones had a copious amount of experience with studying infertility in couples, using endocrinological techniques. At this time, there was no substantial research on endocrinology and the link to infertility so Jones submitted an article of her findings to the American Medical Association titled "Some Newer Aspects of the Management of Infertility". [ 5 ]  Within this article was the advancements she has made studying the luteal phase defect, a term which Jones is responsible for. In 1949, Jones made the first description of  Luteal Phase Dysfunction , and is credited to be the first in using  progesterone  to treat women with a history of  miscarriages , thus allowing many of them to not only conceive, but to deliver healthy babies. [ 6 ]  The pregnanediol technique was developed by Jones along with other Hopkins members. Conclusions drawn from her individual research showed that low progesterone levels lead to low preganediol levels and provide a greater risk for infertility. [ 5 ] [ 7 ]


She became the director of Johns Hopkins' Laboratory of Reproductive Physiology and was the Gynecologist-in-Charge of the hospital's gynecologic endocrinology clinic in 1939. She married  Howard W. Jones  while at Johns Hopkins and they had three children.


In 1969, Seegar Jones began to identify and examine what is now known as ovarian resistance syndrome. [ 7 ]  She demonstrated that stimulation of menopausal gonadotropin leads to the increase in number of eggs that are available and viable for vitro fertilization. [ 8 ]  In 1978, the same year that UK scientists were successful with in vitro fertilization, the Joneses took an opportunity from EVMS and moved to Norfolk, Virginia to create an IVF program in the United States. This was after the birth of the first  test tube baby  in the world,  Louise Joy Brown , on July 25, 1978, in England. [ 1 ]  The Joneses created their own IVF program at EVMS. On December 28, 1981, their procedure gave birth to  Elizabeth Jordan Carr , the first American  test tube baby .


Jones died on March 26, 2005, in  Portsmouth, Virginia . [ 1 ]


Dr. Seegar Jones was awarded the Distinguished Service Award Medal from the Cosmopolitan Club of Norfolk in 1988 for the advancements she made to in vitro fertilization. She is also a recipient of the Dean's Outstanding Faculty Award from Eastern Virginia Medical School, 1996;  and in 1997, the Distinguished Alumnus Award from Johns Hopkins University, also for her contributions to reproductive endocrinology and success she made with IVF. Five years prior to her passing, 2000, she received the Distinguished Scientist Award from the Gynecologic Investigation. [ 9 ]


She was one of the first in her field to train medical students, residents and fellows for several schools. Her dedication to bettering the physician generations below her paved the way of academic medicine. [ 2 ]  One of the many reason why Dr. Seegar Jones is so distinguished and honored in her field. She was named the first woman President of the American Fertility Society in 1970. [ 2 ]  One of the reasons she was honored with this title is because her work with fertility prompted women to control their future.


Sammy Lee  (born  Samuel Lee , 1958 – 21 July 2012) was an expert on  fertility [ 1 ]  and  in vitro fertilisation [ 2 ] [ 3 ]


He was a hospital scientific consultant and was the chief scientist at the Wellington IVF programme. [ 4 ]  His book  Counselling in Male Infertility  was published in 1996; he contributed to major newspaper articles and appeared on several current affairs television programmes. He was the "inspiration" for  Anthony Ling , the character in the novel  One Life  by Rebecca Frayn (Simon & Schuster 2006,  ISBN   0-7432-6876-8 ), after the author herself sought Lee's help for IVF treatment. [ 5 ]


In 2010,  Willing to Die for It , Lee's biography by  Frances Lynn  was published by  Murray Print . [ 6 ]


Lee's interests lay in the field of  stem cell  biology and  regenerative medicine . He was a visiting professor at the  University College London  where he collaborated with various groups in the Anatomy Department examining the potential of  bone marrow  derived  mesenchymal stem cells  to give rise to  neuronal / glial  lineages in response to various  growth factors  and  tissue culture  manipulations. He also taught ethics of biomedicine at  University College London .


In addition, he was interested in tissue engineering and teaching ethics in reproduction. He was attached to the Jessen-Mirsky laboratory and was studying remyelination. Lee had ethical committee approval in Brazil for creating artificial gametes from umbilical cord blood derived stem cells. It is hoped these will be used to overcome infertility. [ 7 ]


Lee originally researched his Ph.D. at UCL under the supervision of Professor  Ricardo Miledi  FRS in the school of  Sir Bernard Katz . [ 8 ] [ failed verification ]  Lee was also a signatory to a memento presented to  Sir Bernard Katz  on his 80th birthday from bona fide members of the school-occasion when the phrase "School of Katz" was coined. [ 9 ]


Lee first published research in  Neuroscience  during the 1980s (from the  Biophysics  department at UCL), having worked on nerve muscle interaction ranging from examining TTX-resistant action potentials in denervated muscle  [ 10 ]  to studying cholinesterase activity in dissociated adult muscle fibres  [ 11 ]  following up on the work first done in the same department by Bill Betz and  Bert Sakmann . [ 12 ] [ 13 ]  Lee changed his focus to embryos, when he realised that many of the questions framed by his  Neuroscience  research were rooted in the matter of differentiation. The ultimate undifferentiated cell is the fertilised egg. This led Lee to work on  gap junctions  in early mammalian embryos (in the Anatomy & Embryology department at UCL), where work with  Anne Warner  FRS and  Anne McLaren  DBE FRS produced new information on factors affecting communication between cells and their developmental potential. [ 14 ] [ 15 ]


Lee became a clinical embryologist in 1985, when working with the gynaecologist  Ian Craft [ 16 ]  he directed the IVF laboratory at the  Wellington Hospital  in  London , then one of the largest units in the world. Consultancy work with the UK division of Ares Serono (1986–1994) also involved work with the Bourn Hallam Group, which  Patrick Steptoe  and  Bob Edwards  had set up after  Louise Brown 's birth. From 1995 to 2002 he was based at the  Portland Hospital  for Women & Children. [ 17 ] [ 18 ]


Lee latterly became based again at  UCL . He was interested in  tissue engineering  and teaching  ethics  in reproduction. [ 19 ]  He was working on tissue engineering and stem cells mainly concerned with background  research  on  Stem cells  and  Progenitor cells . Dr. Lee also had a keen interest in ethics and expressed this at UCL by running a course titled "Ethics of Fertility and Embryo Research". His students remarked him to be knowledgeable in the field of ethics and enthusiastic; they had particularly noted that he is clearly a fan of Kant.


He had previously served as an international  editor  of the Brazilian Journal of Assisted Reproduction. He served as a Trustee and director of several charities. His  curriculum vitae  lists numerous  scientific publications . He was elected as a fellow of the Royal Society of Health (now  Royal Society for Public Health ) and the  Royal Society of Medicine , both in 1987. [ 20 ]


Lee's team at the Wellington pioneered the first UK practise of gamete Intra-fallopian transfer ( GIFT ),  The GIFT of Life  (a technique invented by  Ricardo Asch  in the USA). The team was the second to carry out GIFT and then proceeded to post the largest series in the world in 1986. [ 21 ] [ 22 ]


Lee also helped perform some of the first  egg   donations  in the  UK , when directing the  Wellington Hospital  IVF Laboratory. [ 23 ]


Lee pioneered a simple inexpensive efficient form of Mechanical Assisted Hatching in the  UK . [ citation needed ]  He produced the world's first intra-cytoplasmic sperm injection ( ICSI  with NASBA  virus   assay ) virus free baby to an  HIV  discordant couple. [ 24 ]


Lee had many  media  appearances having been on many  TV shows  as well as writing a number of  newspapers  articles . He wrote several books of his own: such as  Counselling in Male Infertility , and was also a  contributor / consultant  in such books:  The Miraculous World of The Unborn Baby  (Quadrillion, Godalming, Surrey 1998)  ISBN   1-85833-966-9 ,  Natural Fertility  (Hamlyn 2002)  ISBN   0-600-60396-2  and  Inconceivable Conceptions .


Lee was no stranger to  controversy . In addition to his work in IVF, he had differences of opinion with the  HFEA , most notably over the issue of  human cloning . [ 25 ]  He states that if it may be done safely and effectively, and be  morally  justifiable.


Lee also wrote an article in the  Sunday Times  (10 November 2002) in the aftermath of a number of high-profile embryo transfer mixups in several human IVF  clinics .


Professor Sammy Lee arranged a conference co-sponsored by the Progress Educational Trust entitled '21st Century Motherhood' at the University College London (UCL) (18 September 2009).
Speakers included  Peter Brinsden , Consultant Medical Director at Bourn Hall Clinic, Professor Lord  Robert Winston  and Professor  Shere Hite [ 26 ] [ 27 ]
. [ 28 ]


Lee died suddenly on 21 July 2012. [ 3 ]


Lee was educated at  Mill Hill School  and received a  B.Sc.  in  Physiology  in 1979 from  Chelsea College , (later merged into  King's College London ). He received a Ph.D. in Biophysics from  University College London  under the supervision of Prof.  Ricardo Miledi , in a group associated with Sir  Bernard Katz . He later received a  Diploma  in  Counselling  from  London Hospital Medical College  in 1991, and in 2004 a  PGCE  at the  University of Greenwich .


Lee held the following appointments:


James LePage, et al. v. The Center for Reproductive Medicine and Mobile Infirmary Association [ a ]  is a 2024  Alabama Supreme Court  case in which the court reaffirmed that  frozen embryos  are considered a minor child for statutory purposes, allowing for  in vitro fertilization  (IVF) clinics to be held liable for the accidental loss of embryos under Alabama's Wrongful Death of a Minor statute, enacted by the Alabama legislature in 1872. [ 3 ]  In response, several IVF clinics in Alabama suspended operations.


The Alabama Supreme Court's ruling received bipartisan criticism, particularly in the wake of the  U.S. Supreme Court's  decision in  Dobbs v. Jackson Women's Health Organization  (2022).


In vitro fertilization (IVF) is a medical process used to help individuals or couples conceive a child when natural conception is difficult or not possible. It involves combining a woman’s eggs and a man’s sperm in a laboratory to create embryos outside the body. After the embryos develop to a certain stage, typically a few days, one or more are implanted into the woman’s uterus with the hope of achieving pregnancy. The remaining embryos may be frozen for future use, allowing for additional attempts at pregnancy without repeating the entire process.


Between 2013 and 2016, plaintiffs—three married couples—underwent IVF treatment at a clinic operated by the Center for Reproductive Medicine. There, the couples were able to produce several embryos in vitro. Some of those embryos were successfully implanted, and the others were kept in the Center's cryogenic nursery for future implantation. In 2020, a patient at the Center for Reproductive Medicine in  Mobile, Alabama  improperly accessed the cryogenic freezer where frozen embryos were stored, removing and dropping them on the floor after their hands suffered from cold burns. [ 4 ]


Four parents—James and Emily LePage and Felicia and Scott Aysenne—sued against the Center for Reproductive Medicine, with William Tripp and Caroline Fonde serving as plaintiffs in LePage's lawsuit. [ 5 ]  Both suits asserted claims under Alabama's Wrongful Death of a Minor Statute. The Center for Reproductive Medicine moved to dismiss the claims. The trial court granted those motions, explaining its view that "[t]he cryopreserved, in vitro embryos involved in this case did not fit within the statutory definition of 'person'" or "'child,'" and held that their loss did not accrue a  cause of action  for wrongful death. The Mobile Circuit Court affirmed the judgment on appeal. [ 5 ] [ 6 ]


Consistent with a prior decision from 2011, [ 7 ]  on February 20, 2024, the  Alabama Supreme Court  reversed the decision of the Mobile Circuit Court, and remanded the case to the  trial court  for hearing on the liability of the defendants. [ 5 ] [ 8 ]


The Alabama Supreme Court explained in its opinion, with Associate Justice  Jay Mitchell  writing for the majority, that if a frozen embryo is destroyed, those responsible could be sued under the Alabama Wrongful Death of a Minor Statute. [ 6 ] [ 9 ] [ 10 ]  Mitchell's opinion also cited a 2018 amendment to the state constitution, [ 11 ]  arguing that its approval by Alabama voters evidenced "the public policy of [Alabama] to recognize and support the sanctity of unborn life and the rights of unborn children." [ 8 ]


Chief Justice  Tom Parker , elected to his post in 2018, wrote a  concurring opinion  to the ruling, citing the  Bible 's  Book of Jeremiah  and  Book of Genesis ; the  Sixth Commandment ; the  Manhattan Declaration ; as well as Christian  theologians  such as  Thomas Aquinas  and  John Calvin . [ 12 ] [ 13 ] [ 14 ] [ 15 ]  Parker stated:


In summary, the  theologically  based view of the sanctity of life adopted by the People of Alabama encompasses the following: (1) God made every person in His image; (2) each person therefore has a value that far exceeds the ability of human beings to calculate; and (3) human life cannot be wrongfully destroyed without incurring the wrath of a holy God, who views the destruction of His image as an affront to Himself. [ 12 ]


Alabama's Human Life Protection Act, enacted by the Alabama legislature in 2021, which states that it is a felony to perform any type of abortion, was cited by Justice Brady Mendheim when concurring in the court's judgment. [ 15 ] [ 16 ]