L6 key slides meseguer
- 1. Time-Lapse -Cleavage-stage development is a dynamic process in which embryo morphology may change significantly over a time span (h). -Conventional grading practices may not detect subtle differences between individual embryos, such as the time to progress from one cleavage division to the next. -The use of an automated instrument with programmable time-lapse image acquisition, allows data to be collected for individual embryos during development to quantify the exact timing of each cell division.
- 2. 5 15 25 35 45 Day 3Embryo CategoriesASEBIR %Inside %Outside %Inside 33,2 17,1 20,4 29,4 %Outside 35,6 15,7 20,6 28,1 A B C D p=0,941
- 3. Exact timing by time-lapse Precise definition of exact timings (n=9530) Variable Media CI 95% Minimum Maximum Lower limit Upper limit t2 27,93 27,88 28,11 18,40 74,45 t3 38,21 38,07 38,35 19,27 109,45 t4 40,71 40,56 40,85 20,24 94,4 t5 51,03 50,83 51,22 20,29 112,46 t6 54,06 53,88 54,25 25,17 115,6 t7 56,72 56,52 56,92 27,21 116,21 t8 59,11 58,86 59,35 33,37 116,21 Morula 86,63 85,85 87,41 59,95 99,65 Blastocyst 104,12 102,72 104,26 79,94 136,62 ICSI ~ 14:00h D1 17:56h D2 04:13h 06:43h 17:00h 20:00h 22:43h D3 01:07h D4 04:38h 22:07h
- 5. Implantation Success; ESDMorphology included ok Grade A Grade B Grade C Grade D Grade E Discarded non viable excluded yes no yes nono yes A+ A E+ EB+ B C+ C D+ D CC2 5- 12h CC2 5-12h CC2 5-12h CC2 5-12h CC2 5-12h yes no yes no yes no yes no yes no Exclusion Criteria Direct Cleavage Uneven Blastomere T5 48-56h T3 35-40h T3 35-40h embryo kinetics and implantation
- 7. Time-lapse + respiration Respiration Peak = Citokinesis Time-hours post ICSI Embryo respiration and Implantation 5 5,2 5,4 5,6 5,8 6 6,2 6,4 6,6 6,8 7 0 1 2 3 4 5 6 7 8 9 10 fmol/sec Cleavage O2 consumption pattern Not Implanted Implanted
- 8. Cumulus cells — Transcriptomics at an early stage - To identify reliable genomic biomarkers expressed in cumulus cells that accurately and non-invasively can predict the oocyte developmental competence and reinforce the already used morphological criteria. - Gene expression assessed using microarray platforms - The oocyte triggering stimulates the expression of genes involved in the granulosa luteinization process and ovulation finality − a crucial step during follicular development to produce a fully healthy oocyte - Several genes are also differentially associated to successful pregnancy and implantation
- 9. Investigating spent culture medium Pyruvate Glucose Amino acids Sugars Uptake Hormones Lactate Ammonia Enzymes Production Other factors - A noninvasive metabolomic profile of the culture medium surrounding the embryo with an analysis of the components produced or depleted by them can be achieved by different technologies. - Multivariable analysis could be applied and models based on viability index may be used in the embryo selection process.
- 10. Proteomics • Multiple protein analysis of human embryo secretions offers the chance to expand our perspective of a noninvasive quantification of human embryonic viability and even chromosomal normality avoiding any type of manipulation.