Comparison of Cryotip vs

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Presentation transcript:

Comparison of Cryotip vs Comparison of Cryotip vs. Cryotop for mouse and human blastomere vitrification Diana Valbuena, M.D., Ph.D., Maria Eugenia Póo, B.S., Cristobal Aguilar-Gallardo, Ph.D., Sebastian Martinez, Ana Cristina Cobo, Ph.D., Antonio Pellicer, M.D., Ph.D., Carlos Simón, M.D., Ph.D. Fertility and Sterility Volume 97, Issue 1, Pages 209-217 (January 2012) DOI: 10.1016/j.fertnstert.2011.10.008 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Annexin V, caspase 3, and DAPI in blastomeres vitrified with Cryotip. (A) Positive control for annexin V marker. One hour (B), 4 hours (C), and 24 hours (D) alive post-thawed blastomeres. (E) Positive control for caspase 3 marker. One hour (F), 4 hours (G), and 24 hours (H) alive post-thawed blastomeres. Fertility and Sterility 2012 97, 209-217DOI: (10.1016/j.fertnstert.2011.10.008) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Blastomere semithin sections. (A) Mouse blastomere without gelatin. (B) Mouse blastomere covered with 7% gelatin. (C) Human blastomere without gelatin. (D) Human blastomere with 5% gelatin (×100). (E) Survival rate of mouse full embryos vs. zona-free embryos and single blastomeres with and without gelatin (P<.05 by ANOVA). Fertility and Sterility 2012 97, 209-217DOI: (10.1016/j.fertnstert.2011.10.008) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A) Blastomere biopsy from eight-cell embryo on day 3 of development. (B) Thawed blastomere. (C) Divided blastomere 24 hours after thaw (×20). (D) Detail of divided blastomere at ×40. (E) Attached blastomere on day 2 of co-culture with feeder monolayer. (F) Outgrowth on day 5. Fertility and Sterility 2012 97, 209-217DOI: (10.1016/j.fertnstert.2011.10.008) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Ultrastructural study by electron microscopy. (A–C) Fresh blastomere used as control: (A) global view (scale: 10 μm), (B) detail of plasma membrane integrity and cytoplasm, and (C) nuclear and chromatin detail. (D–F) Degenerated blastomere used as control: (D) global view (scale: 10 μm), (E) detail of plasma membrane integrity and cytoplasm, and (F) nuclear and chromatin detail. (G–I) Blastomere vitrified and warmed with Cryotip method: (G) global view (scale: 10 μm), (H) detail of plasma membrane integrity and cytoplasm, and (I) nuclear and chromatin detail. (J–L) Blastomere vitrified and warmed with Cryotop method: (J) global view (scale: 10 μm), (K) detail of plasma membrane integrity and cytoplasm, and (L) nuclear and chromatin detail. Scale of details: 2 μm. Fertility and Sterility 2012 97, 209-217DOI: (10.1016/j.fertnstert.2011.10.008) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions