Assisted Reproductive Technology (ART)

1. Assisted Reproductive Technology (ART) Prepared by Dr. Abdelsalam Talafha Collage of veterinary medicine, JUST American Board Certified, Comparative Veterinary Obstetrics and Gynecology

2. ART • Infertility • Inability to conceive after 1 year of unprotected and regular sexual intercourse • Primary infertility • Couples have never had children • Secondary infertility • Couples initiated conception in the past and then had difficulty

3. ART • USA women infertility rate • Ages 20-24: 4.1% • Ages 25-29: 5.5% • Ages 30-34: 9.4% • Ages 35-39: 19.7% • 80% of infertility cases can be diagnosed • 85% of cases can be successfully treated

4. ART • Female infertility • Disorders of ovulation: 27% • Fallopian tube disorders: 22% • Pelvic adhesions: 12% • Endometriosis: 5% to 15% • Hyperprolactinemia: 7%

5. Male Infertility 100 Total 2104

6. Male Infertility

7. ARTAssisted Conception

8. Assisted Conception • Techniques designed to assist couples in their effort to procreate • Reason • Identification and correction of female, male or a combination of both infertility causes

9. Assisted Conception Objective • To bring sperm and oocyte close to each other to promote chances of fertilization and, ultimately, achieve a pregnancy

11. Assisted Conception Required procedures • Superovulation • Sperm preparation • Assisted fertilization

12. Superovulation • Hormonal manipulation to enhance ovulation and release multiple oocytes during ovulatory cycle

13. Superovulation Drugs used: • Human menopausal gonadotropin • Taken from urine of postmenopausal women • Follicle stimulating hormone (FSH) and luteinizing hormone (LH) activity • Recombinant FSH • Recombinant LH

14. Superovulation- protocol • Gonadotropin for 9-11 days • Monitoring follicular development by transvaginal ultrasound • Follicles 16 -18 mm in diameter • 10,000 IU hCG • Oocytes maturation • Ovulation

16. Sperm Preparation • Select PMNS (Progressively motile normal sperm) • Remove seminal plasma, WBC, and bacteria • Sperm capacitation • Coating of sperm with seminal plasma proteins • Allow sperm to become fertile • In vivo or in test tube

18. Intrauterine Insemination • Sperm sample deposited in uterus just before release of an oocyte (s) in a natural or stimulated cycle using a Soft catheter • Give hCG at injection or up to 24 hrs later • Sperm volume: 0.2-0.3 ml • Pregnancy rates • Around 15% per cycle

19. Gamete Intrafallopian Transfer • Laparoscopic technique in which oocyte and sperm placed in fallopian tube, allowing in vivo fertilization • Procedure • Superovulation • US guided transvaginal oocyte retrieval • 0.1-0.2 mil sperm with 2-3 oocytes

21. In vitro fertilization - IVF • Taking oocytes from woman • Fertilizing them in lab with her partner's sperm • Transferring resulting embryos back to her uterus 3 or 5 days later

22. IVF • Procedure • Superovulation • Insemination • Embryo transfer • Luteal support

23. IVF - Superovulation • Gonadotropin stimulation • Monitoring follicular development • Ultra Sound guided transvaginal oocyte retrieval • Oocyte fertilization with sperm

26. IVF - Insemination • Containers used • Test tubes, Petri dishes, multi-well dishes • Each oocyte inseminated with 0.5-1.0 mil PMNS • Fertilization detected 12-20 hrs later by presence of • 2 pronuclei in oocyte cytoplasm • 2 polar bodies in perivitelline space

27. IVF - Insemination • Syngamy (combination of maternal and paternal pronuclei 24 hrs after insemination) • Further cleavages occur at 24 hr intervals

28. IVF - Embryo transfer • Embryos transferred to uterus on 2nd or 3rd day after in vitro insemination • 4-8 cells embryos • 2-3 embryos transferred in 20 µl of culture fluid • Transabdominal US to see fluid placed in uterus • Cryopreserve excess embryos

29. IVF - Luteal support • Progesterone (P4) necessary for pregnancy maintenance • Premature luteolysis in some superovulatory regimens • P4 supplementation until menses occur or woman has positive pregnancy test

34. Intracytoplasmic Sperm Injection - ICSI • Injection of single sperm into single oocyte in order to get fertilization • Procedure • Superovulation • US guided transvaginal oocyte retrieval • IVF • Oocytes injected with sperm using special microscopes, needles and micromanipulation equipment

40. ICSI - Indications • Low sperm concentration, motility, abnormal morphology • Antisperm antibodies • Fertilization failure after conventional IVF • Ejaculatory disorders • Absence of vas deferens or obstruction of ejaculatory ducts

42. Assisted Hatching Indications • Couples having IVF with • Female partner's age over 37 • Poor quality embryos • Excessive fragmentation • Slow rates of cell division

43. Assisted Hatching – Procedure • Embryo held with a specialized holding pipette • A needle used to expel an acidic solution against ZP • A small hole made in ZP • Embryo washed and put back in culture in incubator • ET shortly after hatching procedure • Hope for the best

47. Further Advances And Uses Of Assisted Conception Technology • Cryopreservation of • Sperm • Embryo • Oocyte • Ovarian tissue • Growth of human follicles and oocytes in vitro • In vitro maturation and transplantation of human spermatozoa

48. Preimplantation Genetic Diagnosis (PGD) • Identify genetic conditions in embryo before ET • Hemophilia • Cystic fibrosis • Aneuploidy

49. PGD Hemophilia • Hereditary bleeding disorder • Absence of a blood protein essential for clotting • Types A: lack of factor VIII • Type B: lack of factor IX

50. PGD Cystic Fbrosis • Genetic disease • Defective gene causes body to produce abnormally thick, sticky mucus that obstruct • Lungs • Pancreas