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Time lapse observations in pre-implantation embryos and factors effecting implantation potential Giles Palmer
ART today ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Andersen (ESHRE) 2009, Seli 2011
At present ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Andersen (ESHRE) 2009, Seli 2011
Choosing the best embryo!
Elder, 2007
Pronuclear Morphology *
Pronuclear Morphology *
Pronuclei Orientation Balaban. Hum. Reprod. 16.11. (2001)
Pronuclei Orientation Current hypothesis: pronuclei to be Parallel to the polar axis
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
2010 Scoring System ,[object Object],[object Object],[object Object],[object Object],Franscovits, 2006
2010 Scoring System ,[object Object],[object Object],[object Object],[object Object],Franscovits, 2006
 
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%)
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%)
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
Cell Cycle Nuclear organization and Mitosis Cellular organization  Cell division Cell synchronicity
Cell Cycle Nuclear organization and Mitosis Cellular organization  Cell division Cell synchronicity Zygotic Clock Maternal to zygotic Γονιμοποίηση Μητρικός έλεγχος Ζυγωτικός  έλεγχος
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)
Time lapse ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Time lapse ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Time lapse
Time lapse ,[object Object],[object Object],[object Object],[object Object],Vajta et al., 2008
 
 
 
 
Time lapse Parameters
 
 
 
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%)
Fragmentation
In vitro study with mouse e m bryos Prediction of developmental potential, Pribenszky, 2010 Importance of the precise detection of early clea v ages
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
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
Importance of fragmentations Duration of the appearing/   disappearing fragments ,[object Object],[object Object],[object Object],[object Object]
1-3 cell division Abnormal Cell division
Abnormal cell division, odd numbers of cells linked to aneuploidy
Time line profile e Time line profile e e CGH profile- molecular karyotype 3-4 Cells
What we have learnt? ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
What we have learnt? ,[object Object],[object Object],[object Object],[object Object]
What we have learnt? ,[object Object],[object Object],[object Object],[object Object]
What we have learnt? ,[object Object],[object Object],[object Object],[object Object]

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Timelapseuk

  • 1. Time lapse observations in pre-implantation embryos and factors effecting implantation potential Giles Palmer
  • 2.
  • 3.
  • 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
  • 11.
  • 12.
  • 13.  
  • 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)
  • 20.
  • 21.
  • 23.
  • 24.  
  • 25.  
  • 26.  
  • 27.  
  • 29.  
  • 30.  
  • 31.  
  • 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%)
  • 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
  • 37.
  • 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
  • 41.
  • 42.
  • 43.
  • 44.

Editor's Notes

  1. Embryo graded 1-5 PN on day1 Cleavage patterns Rate of first cleavage Blastocyst development, cell number, rate, inner cell mass aneuploidy
  2. Embryo graded 1-5 PN on day1 Cleavage patterns Rate of first cleavage Blastocyst development, cell number, rate, inner cell mass aneuploidy