time lapse observations

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time lapse and embryo morphokinetics

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  • 1. Time lapse observations of pre-implantation embryos Giles Palmer, Mitera ACU, Athens gpalmer@mitera.gr

2. There are no commercial relationships or other activities that might be perceived as a potential conflict of interest 3. Time lapse in the IVF Laboratory Continuous viewing of embryo development Embryo monitored from inside incubator Creates archives of embryo development used to select embryo(s) for embryo transfer 4. Embryo selection Embryo selection- subjective assessment Call for consensus of embryo grading Early cleavage, pronuclear morphology & orientation limited Trend to reduce multiple births after ART 5. Embryo selection Invasive techniques Euploid selection (PGS) not fulfilled expectations CGH replacing FISH RCT underway Non-invasive techniques-omics search for biomarkers: metabolomics- not implemented at present. Promising developments but time consuming/ technically challenging 6. The (Short) History of Time lapse monitoring Payne (1997) Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time lapse cinematography Pribenszky (2010) Pregnancy achieved by transfer of a single blastocyst selected by time lapse monitoring Wong (2010) Non-invasive imaging of human embryos before embryonic genome activation predicts development to blastocyst stage Meseguer (2011) The use of morphokinetics as a predictor of embryo implantation 7. Time lapse 2013 Primo Vision (Bright field) Embryoscope (Bright field-self contained incubation) Eeva (Dark field, automatic embryo prediction software) Conventional incubator Time lapse microscope Microwell embryo culture dish with the developing embryos 8. Static observations are misleading! Fragmentation: Pribenszky (2010) 12% embryos fragmenting 89% reabsorbed Average time for fragments to appear/disappear 9.1 h ( +/- 442) Chance for NOT noticing fragments at bi-daily monitoring: 72% !!! Blastocyst contractions: 9. videos Which embryo would you transfer? Static observations are misleading! 10. Ww 26:32 29:52 30:02 35:12 42:02 42:32 11. Implantation is linked to exact timing events Event PN appear PN fading* 1st division* 2nd division* 3rd 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%) Herrero, 2010 12. 5-8cc 30-50 min 9-16cc 40-70 min 3-4cc 10-20 min 8-9cc 22-24 hrs 4-5cc 14-16 hrs 2-3cc 10-12 hrs Interphase 1-2cc 20-26 hrs Exact timing of interphases 1st cycle 2nd cycle 3rd cycle 4th cycle Hlinka 2010 13. Cell Cycle Cellular organization Nuclear organization and Mitosis Cell division Cell synchronicity Karyokinetics and cytokinetics 14. Karyokinetics and cytokinetics 15. Karyokinetics and cytokinetics Second cell cycle Third cell cycle 3-4 cell 5-8 cell 16. t2 t3 t4 t5 cc2 cc3s2 t: exact time t2: time to 2-cell t3: time to 4-cell t4: time to 4-cell t5: time to 5-cell cc: cell cycle cc2 = t3-t2 cc3 = t5-t4 S:synchrony s2 = t4-t3 cc2=11,8h s2 =