Assisted/Advanced Reproductive Technology

1. Assisted/Advanced Reproductive Technology Tyler Dohlman

2. Outline • 1st lecture • ART technologies • Artificial/Intrauterine Insemination • Embryo transfer • IVF • Cloning • 2nd lecture • ART problems • Cloning • IVF humans

3. Possibilities • Artificial Insemination • Embryo Transfer • In vitro Fertilization • Trans vaginal oocyte recovery (TVOR) • Intracytoplasmic Sperm Injection (ICSI) • Zygote Intrafallopian Transfer (ZIFT) • Gamete Intrafallopian Transfer (GIFT) • Cloning (Somatic Cell Nuclear Transfer) SCNT • Embryo splitting • Gene transfer

4. Artificial Insemination • 1st advanced reproductive technique (1784-dog) Spallanzani • Today = about every animal known to man has used this technique to advance genetic production or to conserve genetics • Production animals vs companion animals vs zoo (extinction species) vs humans conception

5. AI Technology • Collecting sperm cells through live collection or artificially • Using fresh, chilled, or frozen semen • Most semen products are extended with solutions to preserve integrity of cells, nutrients are added for cells to stay alive, and cryoprotective agents are also added for freezing, with addition of antibiotics to kill opportunistic pathogens

6. AI Techniques • Intra/trans cervical insemination • Intrauterine insemination • Surgical • Non-surgical • Laproscopic • Intratubal insemination • Intrauterine tuboperitoneal insemination

7. Animal Data (AI) • National Animal Health Monitoring System • Survey data • Beef cattle = 8% (2007-2008) • Dairy Cattle = 75% (2007) {~72.5% of all pregnancies} • Swine = 85% (2000) • Poultry = unknown but it is A LOT! • Equine = unknown, registration only

8. AI Human Data • The first reported case of artificial insemination was in Philadelphia (human 1884) but was not reported for 25 years -"Letter to the Editor: Artificial Impregnation"The Medical World: 163–164. April 1909.  (cited in Gregoire, A. and Mayer, R. (1964). "The impregnators". Fertility and Sterility (16): 130–134. ) • In 1987, >170 000 women in the USA were treated for infertility using artificial insemination (Critser, 1998) • Approximately 3000 couples seek AID treatment annually in the UK, and there were >16 000 treatment cycles in 1992 (Human Fertilisation and Embryology Authority, 1994)

9. Human AI (IUI) • Reasons • Male issues • Unexplained infertility • Low sperm count • Decreased sperm mobility • Ejaculation dysfunction • Requirement of donor sperm • Female issues • A hostile cervical condition, such as cervical mucus that is too thick • Cervical scar tissue from past procedures or endometriosis

10. Statistics • ~680,000 females received IUI in 2002 • Average ~746,000 females received IUI (2006-2010)

11. IUI Study • “Results of 6139 artificial insemination cycles with donor spermatozoa” • Cumulative pregnancy rate (PR) for 3, 6 and 12 months were 36, 53 and 75% respectively. • Cumulative PR (age groups) • <37 years: 44, 61 and 82% • 37–40: 24, 44 and 61% • >40 years: 13, 22 and 50% • Botchan et al. Hum. Reprod. (2001) 16 (11): 2298-2304. doi: 10.1093/humrep/16.11.2298

12. IUI Trend

13. IUI 2nd study • “Infertility: Prognostic indicators for intrauterine insemination (IUI): statistical model for IUI success” • A retrospective analysis of 260 completed intrauterine insemination (IUI) (Male factor problems were excluded) • Overall pregnancy rate = 19.6% per completed cycle • Miscarriage rate 15.6% • Multiple pregnancy rate 23.5% • Cancellation rate 19% • 4 significant IUI variables • follicle number (P < 0.005) • chance of conceiving : one follicle = 7.6%, two follicles = increased to 26%. • endometrial thickness (P < 0.005) • duration of infertility (P < 0.01) • progressive motility (P < 0.05)] • Tomlinson et al. Hum. Reprod. (1996) 11 (9): 1892-1896.

14. Embryo Transfer • 1st successful embryo transfer (1890 – rabbit) • 1st bovine embryo transfer (1950-Wisconsin) • 1st human successful transfer (1983-UCLA) • Blakeslee, Sandra (1984-02-04). "Infertile Woman Has Baby Through Embryo Transfer“. The New York Times. Retrieved 2010-05-01.

15. Embryo Transfer Techniques • Surgical • Non-surgical http://www.fao.org/docrep/004/t0120e/t0120e07.htm

16. Bovine Embryo Transfer • http://www.fao.org/docrep/004/t0120e/T0120E05.htm

19. Human Embryo Transfer • >90% of embryos are put in via IVF

20. In vitro Fertilization • The first attempt at IVF on mammalian(rabbit, guinea pig) eggs (Viennese embryologist Samuel Leopold Schenk -1878) • 1951 – (Colin Russell Austin in Australia and Min Chueh Chang in the United Sates) figured out spermatozoa needed to have capacity to fertilize “capacitation” • 1959 - Chang successfully use IVF to impregnate a rabbit • First successful fertilization of aspirated human ova (1944-Rock and Menkin) • The first successful birth from IVF (non stimulated) “Louise Brown” in 1978. Dr. Robert Edwards, the physiologist who developed the treatment, was awarded the Nobel Prize in Physiology or Medicine in 2010.

21. IVF Technique In Vitro Fertilization • http://www.ib.bioninja.com.au/standard-level/topic-6-human-health-and/66-reproduction.html

22. IVF Cattle • Results vary • Average 18 oocytes/aspiration • 30 % develop into a viable embryo • Average 4.9 transferrable (Grade 1 & 2) embryos per IVF cycle • 40-50% conception rates • http://www.transova.com/our-services/ivf.php • With conventional ET and IVF = >1.1 billion embryos produced each year in the world

25. IVF Human • Most common ART in human medicine • >147,000 IVF cycles done a year in US • In 2010, >61,000 births from IVF in US (1-2% all births)

27. IVF Human (CDC annual report)

28. Intracytoplasmic Sperm Injection • https://www.youtube.com/watch?v=IQujLI-ArMY

29. IVF Success

30. Advancements in IVF • Anti-Mullerian Hormone (AMH) • Produced by granulosa cells of ovary • Highest production is by the primordial to antral follicles <4mm • The higher the AMH production = the higher follicular pool/reserve • IVF clinics have started using this as a primary marker if IVF will be pursued and what success whould be expected • Has been used in cattle IVF and Embryo production systems

31. http://www.rmanj.com/2013/09/anti-mullerian-hormone-amh-testing-of-ovarian-reserve/http://www.rmanj.com/2013/09/anti-mullerian-hormone-amh-testing-of-ovarian-reserve/

32. Advancements in IVF • Genomics in cattle • Greater accuracy of predicting genetic merit of young animals • Eliminating time in generation interval to advance genetic merit • Increase intensity in selection of superior genetic animals at a young age • Intense selection of certain traits (production vs health vs conformation) • Other uses: parenting testing, identifying deleterious recessive traits or defects

33. Cloning (Somatic Cell Nuclear Transfer) • In 1928, Hans Spemann,(German embryologist) and Hilde Mangold had 1st somatic-cell nuclear transfer (amphibian embryos) • “Dolly”- 1st mammal cloned • Mammary tissue “donor cell” • Megan and Morag June 1995

34. SCNT Cloning Technique • Transfer nucleus from a donor adult cell (somatic cell) to an enucleated egg http://en.wikipedia.org/wiki/File:Dolly_clone.svg

35. Cloning Cattle • Viagen (TransOva) • Preserving genetic • Pigs, cattle, small ruminants, horses, and pets

36. Cloning (Human) • 2 types • Therapeutic cloning and reproductive cloning • Therapeutic = creating stem cells for medicinal purposes and research • Reproductive = creating cloned human • 1st hybrid human clone (1998-Advanced Cell Technologies) • Donor cell = man's leg cell • Recipient cell = cow's egg • Destroyed after 12 days "Details of hybrid clone revealed". BBC News. June 18, 1999. Retrieved April 30, 2010.

37. Cloning • Legal, moral, and social issues • Is it right or wrong to use the technology for something and not for other purposes? http://globaldiscussion.net/topic/5014-the-pros-and-cons-of-human-cloning/

38. Questions