Mouse Oocytes and Embryos Cryotop-vitrification Using Low Concentrated Solutions: Effects on Meiotic Spindle, Genetic Material Array and Developmental Ability

Objective(s): The examination of the possibility of applying lower CPA- concentrations and obtaining the similar results to those using higher concentrations; as it is shown, the toxicity of the CPAs used in vitrification approach will diminish. Materials and Methods: Following vitrification/warming, oocytes were subjected to PZD/ICSI. SRs, FRs, and DRs were recorded. SRs and DRs of the embryos were monitored after vitrification/warming. IHC studies were done. Data were analyzed in comparison to the data of Exp. (experimental groups) applying 1.5 M CPA- concentrations (largely-used concentration). Results: The data of oocytes exposed to 1.25 M concentrated CPAs were in consistency with those exposed to 1.5 M and fresh oocytes in terms of SRs, FRs and DRs. Normal spindle and chromatin configuration is in consistence between the two experimental groups, but lower in comparison with control group. The lower the concentrations were, the less SRs, FRs, DRs were. Also, spindle organizations were more normal in comparison with the experimental groups as the concentrations decreased. The results of DRs for embryos which were exposed to 1.25 and 1.0 M concentrated CPAs were close to those vitrified with 1.5 M and fresh embryos but IHC observations in the three Exp. were significantly lower than those of fresh embryos. The results of 7.5 M concentrated CPAs solutions were significantly lower than those of the control group 1.5, 1.25 and 1.0 M treated. Conclusions: Vitrification by cryotop technology using minimal volume approach increases both cooling and warming rates, therefore, the CPAs limited reduction to 1.25 and 1.0 M instead of using 1.5 M for oocytes and embryos cryotop-vitrification procedure, may be a slight adjustment.