2020. 11. 02. UNIVERSITY OF VETERINARY MEDICINE, BUDAPEST In vitro fertilization – test tube baby program in animals and man László Solti Professor emeritus 1 Brackett with Willy and Nilly, the first two test tube goats born in the US 2 1 2020. 11. 02. Timeline of in vitro fertilization 3 Embryo development (IVF-ET) 4 2 2020. 11. 02. In vitro fertilization Large Offspring Syndrome Liver, kidney and heart can be affected Common in ruminants (dystocia → †) In vitro culture? 5 Major steps of IVF 6 3 2020. 11. 02. Ultrasound guided Ovum Pick Up (OPU) 7 Instruments and drugs used for IVF 8 4 2020. 11. 02. Hormonal Stimulation for IVF Weekly scheme and hormone treatment for Ovum pick- Blastocyst yields from the animals subjected to OPU up (OPU). (a) The animals were not treated with without FSH (open bars) and OPU with FSH treatment hormone FSH. Transvaginal OPUs (triangle) were (solid bars). performed on Monday and Thursday of each week for consecutive two weeks, resulting in a total of four sessions (Presicce et al, Vet Med Int. 2011; 2011: 145626) (b) FSH treatment: FSH (arrow) at dosage of 80mg IM and 120 SC was simultaneously administered on Day 1 (Saturday). Ovum pick-ups were performed 48 and 96 hours post FSH injection, meaning on Day 3 and Day 5 of each week for consecutive two weeks, resulting in a total of four sessions. 9 Donor preparation (Luis Ferré, INTA Rafaela) 10 5 2020. 11. 02. Follicle aspiration 11 Isolation and maturation of oocytes (Luis Ferré, INTA Rafaela) 12 6 2020. 11. 02. Cumulus-Oocyte-Complex (COC) 13 IVF 14 7 2020. 11. 02. Cumulus-Oocyte-Complex and oocytes at the end of maturation At the end of maturation (Katherine Hendrics) After treatment with hyaluronidase (Firdous Khan) 15 In vitro fertilization and culture (Luis Ferré, INTA Rafaela) 16 8 2020. 11. 02. ET and pregnancy diagnosis (Luis Ferré, INTA Rafaela) 17 Embryo production in vitro Human Animals (ruminants) IVM ø + IVF + + IVC ++ Intermediate recipients ø Cocultivation , ø ++ Ultrasound guided ovum pick up (OPU) + , ø (Human: Edwards and Steptoe 1978 (Louise Brown) Cattle: Brackett et al. 1981, goat: Brackett et al, 1983 18 9 2020. 11. 02. Sperm preparation for IVF (Swim up) Heparin containing medium (M 199) 19 Sperm preparation for IVF Percoll gradient centrifugation 20 10 2020. 11. 02. Sperm preparation for IVF Glass wool filtration 21 MOET vs IVF Parameters MOET IVF Collection/year 6a 24b Embryos/collection 6 4.5 Annual Embryo Production 36 108 Pregnancy Rate (%) 60c 45c TOTAL Born Animals 22 49 a Collections every 60 days b 2 OPUs/month c Fresh transfers (Luis Ferré, INTA Rafaela) 22 11 2020. 11. 02. MOET vs IVF with sexed semen Parameters MOET IVF (sexed semen) Collections/year 6a 24b Embryos/collection 6 3 Annual Embryo Production 36 60 Sex Ratio (Females:Males) 50:50 90:10 Pregnancy Rate (%) 60c 45c TOTAL Females Born 11 29 a Collections every 60 days b 2 OPUs/month c Fresh transfers (Luis Ferré, INTA Rafaela) 23 24 12 2020. 11. 02. 25 ET vs IVF worldwide Source: 27th Annual Report of IETS (2018) 26 13 2020. 11. 02. Production of bovine embryos in Brazil embryos of Number ET: conventional embryo transfer, IVF: embryos produced in a laboratory 27 Source: L.C. Cruz and A. Sarabia ([email protected]) 28 14 2020. 11. 02. Current status of ET and IVF using Ovum Pick Up in cattle 29 Az anyák átlagos életkora első gyermekük születésekor 1990 23 1995 23,5 1998 24,2 2000 25 2005 27 2008 28 0 5 10 15 20 25 30 30 15 2020. 11. 02. In vitro production of human embryos 31 Brave New World is a dystopian novel written in 1931 by English author Aldous Huxley, and published in 1932. Largely set in a futuristic World State of genetically modified citizens and an intelligence-based social hierarchy, the novel anticipates huge scientific developments in reproductive technology, sleep-learning, psychological manipulation, and classical conditioning that are combined to make a utopian society that goes challenged only by a single outsider. (1931) 32 16 2020. 11. 02. "for the development of in vitro fertilization." https://www.youtube.com/watch?v=fy61qs_tXTQ https://www.youtube.com/watch?v=1zQhwiGhzJk https://www.youtube.com/watch?v=TIRbsqvjuRs 33 Births and deaths in Hungary 34 17 2020. 11. 02. World population according to continents, 1950-2100 UN, World Population Prospects 2019 https://www.ksh.hu/interaktiv/grafikonok/vilag_nepessege.html 35 Population data Hungary, June 2017 Live births: 73.300 Deaths: 92.200 Population decrease: 1.900 https://www.ksh.hu/docs/hun/xftp/gyor/nep/nep1706.html 36 18 2020. 11. 02. Decreasing population Natural Population Natural Total fertility Livebirths Deaths increase (in thousands) increase index (per 1000 head) 1960 9 984 146 461 101 525 44 936 4.5 2,02 2005 10 088 97 496 135 732 -38 236 -3.8 1,32 2018 9 778 89 807 131 045 -26 900 -4.2 1,50 37 Population forecast until 2100 Average fertility rate in Hungary is 1.4 which cannot stop the population decline. According to UN the population decreases to approx. 6.5 m by 2100 even if the fertility rate goes up to 1.8 which is the moderate rate. Migration balance is also negative – if it would go down to zero, at moderate fertility rate the population would be below 6 m. An increasing trend to become above 10 m could be reached by a fertility rate >2.3 which is almost impossible. 38 19 2020. 11. 02. Composition of the Hungarian population Arrow: missing age group as a consequence of WW-I 39 Population pyramid 1960-2014 40 20 2020. 11. 02. Patrick Steptoe Bob Edwards • 1970-ies: millions of couples failed to get child. With oviductal patency failure (approx 20%) there was no chance for conception • Lesley and John Brown visited different physicians for 9 years before they came to Dr. Patrick Steptoe (†1988), who performed in vitro fertilization (IVF) in 1977 when it was in experimental phase • July 25, 1978: birth of Louise Brown, the first test tube baby • Since then >8 million IVF children were born worldwide. Pregnancy rates now steady at around 36 percent, according to ESHRE monitoring (Science Daily July 2018) • In spite of resistance of churches, governments and media as well as the scepticism of scientific world, Edwards and Steptoe went on working 41 Patrick Steptoe and Robert Edwards: Brave New Baby Doctors „The astonishing feat brought instant celebrity to Patrick Steptoe and his partner, Robert Edwards - as well as a barrage of criticism. The Roman Catholic Church and other religious groups denounced it as playing God, and even scientists like James Watson, unraveler of DNA, were worried about tinkering with a process as sacrosanct as procreation. But the debate faded as it became clear that the brave new world of babymaking that Steptoe had ushered in was providing a desperately sought service.” 42 21 2020. 11. 02. • The history of IVF and ET dates back as early as the 1890's when Walter Heape successfully transferred embryos between rabbits, long before the applications to human fertility were even suggested • „We were called everything under the sun - immoral, unethical, dehumanising” 43 BBC News ( 10/4/2010) Robert Edwards, pioneer of in vitro fertilization (IVF), has won this year's Nobel Prize in medicine, and the award carries a resonance in modern science Livemint ( 10/4/2010) Very few scientists can say that four million people are alive because of their work, but Robert Edwards is one of those few. ZDNet (blog) ( 10/4/2010) A Vatican official has said the awarding of the Nobel Prize for Medicine to British IVF pioneer Robert Edwards is "completely out of order" National Geographic (10/4/2010) Vatican bioethics official: Nobel prize winner started new chapter but IVF not Hindustan Times (10/4/2010) Edwards' prize a political act. If having a family does not require sex, and all that entails, if humanity is something that can come 44 22 2020. 11. 02. 2014 Annual Report, Society for ART (USA) • Since 2007, the birth rate has been declining steadily • 2,000 more IVF babies were born in 2012 than 2011 The average cost of one IVF treatment in the US is $12,400, and that's without the extra medicines the couple may need. Often, couples will need more than one treatment to conceive. The number of multiple embryos being transferred in IVF treatments has also declined • IVF success rates decrease as women age The IVF success rate was significantly higher for women under the age of 35 compared with those who were older. Looking only at the IVF treatments that used fresh embryos from non- donor eggs, the number of women under the age of 35 who gave birth using IVF was about 40%, compared with only 31% for women between the ages of 35 and 37. Only 3.9% of 42-year-olds got pregnant 45 Severino Antinori • Starting in veterinary medicine, he changed to human medicine at the La Sapienza University. Graduated in 1972 • First he worked in gastroenterology but after meeting with de Steptoe changed to gynecology and obstetrics • Main field of interest was infertility, in 1982 he opened his own clinic • In 1989 he extended the IVF activity to elder menopausal women • In 1994 his 63-year-old patient, Rosanna Della Cortes became the oldest pregnant woman giving birth to a sound normal child 46 23 2020. 11. 02. Sunday, 25 July 1993 • Severino Antinori, the Italian obstetrician at the centre of the storm over fertility treatments for older women, is very angry.