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In-vitro maturation: patient selection and results

In-vitro maturation: patient selection and results. Aygul Demirol Assoc Prof, Medical Director GURGAN CLINIC IVF Center , Ankara- Turkey. In-vitro maturation (IVM). Immature oocyte retrieval and subsequent oocyte maturation in vitro without any ovarian stimulation.

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In-vitro maturation: patient selection and results

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  1. In-vitro maturation: patient selection and results AygulDemirol Assoc Prof, MedicalDirector GURGAN CLINIC IVF Center, Ankara-Turkey

  2. In-vitro maturation (IVM) • Immature oocyte retrieval and subsequent oocyte maturation in vitro without any ovarian stimulation

  3. IVM (in-vitro maturation) • In 1991, Cha et al. reported a pregnancy from IVF with oocytes obtained from ovariectomy specimens and matured in culture (healthy triplet girls) (Cha KY, Fertil Steril 1991)

  4. IVM (in-vitro maturation) • Trounson et al. reported the birth of a normal baby with IVM of immature oocytes from a polycystic ovary syndrome (Trounson Fertil Steril 1994)

  5. IVM (in-vitro maturation) • There have been more than 1000 births of babies with IVM procedures, including in patients with PCOS (Chian RC) • BUT, IVM has not become mainstream in IVF, with ovulation induction cycles with oocyte retrieval of mature (MII) oocytes still the highly favored protocol.

  6. Why IVM ? • Reduced cost • Avoiding OHSS • Simplification of treatment compared with conventional IVF-ET

  7. IVM Indications • PCOS • High responders • IVF/IVM • Fertility preservation • Donation cycles • Delayed responders • Male infertility • Poor responders

  8. 2-12 mm follicles

  9. In general • clinical pregnancy rate 30-35% • implantation rate 10-15% (R.C. Chian RBM Online, 2004)

  10. IVM low implantation rates when compared to conventional stimulated cycles. asynchrony in the cytoplasmic and nuclear maturation of the oocyte asynchrony in the endometrium culture conditions

  11. IVM is based on the treatment of • Two main groups of patients • With PCO (have irregular, mostly anovulatory cycles and are at increased risk for OHSS because of their higher sensitivity to gonadotropins) • With normal ovaries (may wish to avoid the side-effects of hormone injections)

  12. How to maximize IVM results byoptimizing clinical management • Patient selection criteria-best candidates • Under 35 years of age • PCO/PCOS

  13. Patient management-I Baseline TV-USG (day 2, 3) Second USG (between day 6 and day 9) (for follicular and endometrial assessment) HCG priming 36 hours prior to egg collection

  14. Patient management-II • IVM ovum aspiration needle, single or double lumen, 19 G, 35 cm • Aspiration pressure 85-100 mmHg • All visible follicles are aspirated

  15. Patient management-III • Priming with FSH or HMG ? • Priming with HCG ?

  16. FSH Priming • Results are conflicting • Potential benefits: • Larger ovarian size • Easier retrieval • Higher E2 levels • More maturational competence May lead to improved endometrial priming

  17. HCG Priming • Theoretically HCG priming • Promote invitro maturation • Improve pregnancy rates However the exact mechanism of HCG on small follicles is still unclear

  18. HCG Priming • First prospective study Chian et al. (2000) • Increased oocyte maturation • High clinical pregnancy rate • ( 36 %)

  19. hCGPriming • In-vitromaturation rate is faster in oocytesobtainedfromhCGprimed IVM oocytes. (Chian et al.,HumanReprod ,2000 ; Son et al., RBM Online,2006) - Hastensthetheoocytematuration in-vitro - Makestheoocyteretrievaleasier • Matureoocyte on theday of retrieval is higher (Son et al.,Hum.Reprod,2002) • Higherfertilization,cleavageandblastocytdevelopmentrates in IVM cycles • Number of goodqualityblastocystshigher (40% vs 23.3%) (Son et al.,RBM Online,2008)

  20. Lab management-I • Determination of cumulus-oocyte complexes (COCs) (special sliding technique-after using cell strainer) • The immature COCs are incubated in culture dish containing 1 ml oocyte maturation medium supplemented with a final concentration of 75 mIU/ml FSH and 75 mIU/ml LH • for SAGE medium

  21. Lab management-II • For MediCult medium, preincubation in LAG medium (2-3 hours) • Transferring into IVM final maturation medium (9 ml IVM medium is added 1 ml patient serum, 10 µl pregnyl, 100 µl FSH)

  22. Lab management-III • Stripping oocytes 24 hours after culture • Twenty for hours after maturation additional 24 hours for immature COCs • Mature oocytes are subjected to ICSI • Embryo maintenance medium for SAGE • ISM1 for MediCult

  23. Endometrial priming • 17-β-oestradiol starts on the day of OPU (2 mg orally, three times daily and continue until pregnancy test) • Two days after OPU, intravaginal progesterone suppositories 600-800 mg, daily and continue until pregnancy test)

  24. Clinical outcome for PCO/PCOS Jurema MW. Fertil Steril 2006;86:1277–91.

  25. Clinical outcome for normal ovaries Jurema MW. Fertil Steril 2006;86:1277–91.

  26. Pregnancies and deliveries after transfer of humanblastocysts derived from in vitro matured oocytes inIVM ( PCO(S) ) (Blastocyst vs cleavage ET)

  27. The abortion rate, gestational age and birth weight at delivery, and obstetric complications of pregnancies conceived by IVM-ET in women with PCOS were comparable with those of other women with PCOS being treated by conventional IVF-ET (Fertil Steril, 2005)

  28. CONCLUSION: Compared with IVF and ICSI, IVM is not associated with any additional risk. (Obstet Gynecol 2007;110:885–91)

  29. Outcome of the IVM cycles (sept 2005-jan 2010) GURGAN CLINIC IVF Center Ankara, Turkey

  30. n % mean Mean age 30.2 Cycles 321 Cycles with oocytes 321 100 Oocytes retrieved 2725 8.7 24 h maturation Oocytes reaching MII 1253 47.1 4.2 Oocytes fertilized(2PN) 989 79.2 3.08 Embryos cleaved 890 90.1 48 h maturation Oocytes reaching MII 408 15.3 1.2 Oocytes fertilized (2PN) 301 74 Embryos cleaved 210 70.2

  31. n % mean transfer and outcome Cycles with embryo transfer 288 90.4 Embryos transferred 806 2.8 Biochemical pregnancies/transfer 103 36.1 Clinical pregnancies/transfer 78 27.1 Implantation rate 13.4 Ongoing pregnancies 41 Live birth 16 Abortion rate 21 26.9

  32. Immature oocyte after oocyte pick-up

  33. Mature oocytes after denudation & ICSI

  34. Fertilised (2 PN)oocytes

  35. Cleavage stage (Day 2) embryos

  36. Blastocyst stage (Day5)

  37. PREGNANCIES IN TURKEY FOLLOWING IN VITRO OOCTYE MATURATION Aygul Demirol, Tamer Sari, Bagdagul Girgin, Erkin Kent, Suleyman Guven, Timur Gurgan 2007, GORM *Two women with history of infertility and PCOS underwent in vitro maturation (IVM) program without controlled ovarian hyperstimulation. The patients were primed with 10.000 IU HCG 36 h before oocyte retrieval. Oocytes-cumulus masses were matured in IVM medium. The matured oocytes were fertilized by ICSI and embryo transfer was performed on day 3

  38. accepted

  39. 10,000 IU HCG + Immature oocyte retriaval IVM instead of cancellation Leading follicle = 12-14 mm IVM for a Second Chance • Recent reports: Risk of OHSS 47 % CLINICAL PREGNANCY No OHSS Lim et al. Fertil Steril 2002

  40. Natural Cycle + IVM But natural cycle yields only 1 single follicle For other non dominant follicles Because IVM is possible even if the dominant follicle is selected Thornton 1998 Fertil Steril IVM may be an option

  41. Thank you for their help and support • Ri-ChengChian, McGillReproductiveCenter, Montreal, Canada • Mette Munk, Jyllinge, Denmark • andothers…

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