Morphokinetic studies on in vitro embryo developoment-choosing the best embryo

1. Time lapse observations in pre-implantation embryos and factors effecting implantation potential Giles Palmer

4. Choosing the best embryo!

5. Elder, 2007

6. Pronuclear Morphology *

7. Pronuclear Morphology *

8. Pronuclei Orientation Balaban. Hum. Reprod. 16.11. (2001)

9. Pronuclei Orientation Current hypothesis: pronuclei to be Parallel to the polar axis

10. Different strategies of day of embryo transfer Day 5 Only at blastocyst stage (Higher implantation) Flexible Day 2-3 or 5 What ever the approach- it relies on morphology- difficult to predict 51% Day 5 embryos preselected on day 3 Rijenders. Hum. Reprod. 13.8. (1998 ) Early cleavage

14. Implantation linked to exact timing events Herrero, 2010 Event PN appear PN fading 1 st division 2 nd division 3 rd division Range (h) 7.8-11.1 Out of range 22.3-25.8 Out of range 24.4-28.2 Out of range 35.3-40.6 Out of range 36.0-41.6 Out of range 100% Implanted N (%) 15 (54%) 13 (46%) 23 (66%) 12 (34%) 23 (66%) 12 (34%) 13 (72%) 5 (28%) 19 (73%) 7 (27%) 0% Implanted N (%) 60 (49%) 62% (51%) 55 (45%) 67 (55%) 57 (46%) 67 (54%) 43 (45%) 52 (55%) 45 (45%) 56 (55%)

15. Implantation linked to exact timing events Herrero, 2010 Event PN appear PN fading * 1 st division * 2 nd division * 3 rd division * Range (h) 7.8-11.1 Out of range 22.3-25.8 Out of range 24.4-28.2 Out of range 35.3-40.6 Out of range 36.0-41.6 Out of range 100% Implanted N (%) 15 (54%) 13 (46%) 23 (66%) 12 (34%) 23 (66%) 12 (34%) 13 (72%) 5 (28%) 19 (73%) 7 (27%) 0% Implanted N (%) 60 (49%) 62% (51%) 55 (45%) 67 (55%) 57 (46%) 67 (54%) 43 (45%) 52 (55%) 45 (45%) 56 (55%)

16. Exact timing of interphases ID3 5-8cc 30-50 min ID4 9-16cc 40-70 min ID2 3-4cc 10-20 min IP4 8-9cc 22-24 hrs IP3 4-5cc 14-16 hrs IP2 2-3cc 10-12 hrs IP1 1-2cc 20-26 hrs 1 st cycle 2 nd cycle 3 rd cycle 4 th cycle Hlinka 2010

17. Cell Cycle Nuclear organization and Mitosis Cellular organization Cell division Cell synchronicity

18. Cell Cycle Nuclear organization and Mitosis Cellular organization Cell division Cell synchronicity Zygotic Clock Maternal to zygotic Γονιμοποίηση Μητρικός έλεγχος Ζυγωτικός έλεγχος

19. t2 t3 t4 t5 cc2 cc3 s2 Cc: cell cycle cc2 = t3-t2 cc3 = t5-t4 s2 = t4-t3 cc2=11,8h s2 = <0.76h t2=25,6h (24,3-25,8h) t3=37,4h (35,4-37,8h) t4=38h (36,4-38,9h) t5=52,3h (48,8-56,6,h)

22. Time lapse

28. Time lapse Parameters

32. ΜΗΤΕΡΑ _ IVF Results 15 cx, 32-42 yrs., ICSI, Day 2-3 ET Pronuclear appearance/disappearance, cleavage patterns, fragmentation Number Pos B-hCG Total Number of embryo transferred Number of embryos implanted 15 8 (53%) 39 12 (31%)

33. Fragmentation

34. In vitro study with mouse e m bryos Prediction of developmental potential, Pribenszky, 2010 Importance of the precise detection of early clea v ages

35. Reduction of probability of reaching the blastocyst stage if a fragmentation occurs during the embryo development In vitro study with mouse e m bryos Prediction of developmental potential, Pribenszky, 2010

36. Reduction of probability of reaching the blastocyst stage if a fragmentation occurs during the embryo development In vitro study with mouse e m bryos Prediction of developmental potential, Pribenszky, 2010

38. 1-3 cell division Abnormal Cell division

39. Abnormal cell division, odd numbers of cells linked to aneuploidy

40. Time line profile e Time line profile e e CGH profile- molecular karyotype 3-4 Cells