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양정숙,배인하,Yang, Jung-Sook,Bae, In-Ha 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3
Objective: This study was to determine the effect of different concentration of calcium III medium on the preimplantational development of zygotes and early 2 cell embryos. Methods: Female mice of ICR strain ($5{\sim}8$ weeks old) were superovulated and mated with fertile males. Zygotes or early 2-cell embryos were collected by flushing the oviducts $31{\sim}32$ hours after hCG injection. The embryos were cultured in various concentrations of $Ca^{2+}$ in medium or with EDTA, EGTA and $Ni^{2+}$. Result and Conclusion: Treatment of high concentraion of $Ca^{2+}$ (3.42 mM $(2X){\sim}17.l$ mM (10X)) in medium didn't develop well compared to the control. Low concentrations of $Ca^{2+}$ (0.214mM $(1/8X){\sim}0.855$ mM (1/2X) were deterimental to development beyond 2-cell stage. EDTA, $Ca^{2+}$ chelating agent was treated with ranged concentrations of EDTA (0.014 $mM{\sim}0.107$ mM) to medium contaning 1.71 mM $Ca^{2+}$ showed beneficial effect to development to blastocyst compared to the control. EGTA, extracellular $Ca^{2+}$ chelator, was treated with ranged concentrations of EGTA ($0.014{\sim}0.107$ mM) to the medium contaning 1.71 mM $Ca^{2+}$. There is no significant difference with the control. $Ni^{2+}$ (50 ${\mu}M$), T-type $Ca^{2+}$-channel blocker was treated to medium contaning low concentration of $Ca^{2+}$. It overcame 2-cell block significantly. Rate of degenerated embryos decreased and developmental rate to morula and blastocyst increased more than low $Ca^{2+}$ concentration alone. Further studies are needed for the overcoming effect of 2-cell block by $Ni^{2+}$.
양정숙,손철,배인하,Yang, Jung-Sook,Sohn, Cherl,Bae, In-Ha 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3
Objective: The study was performed to compare the survival rate and the development of day 2 mouse embryos which had freezing procedures done. Methods: We used three different vitrification solutions (EFS, VS14, DPS) and a ultrarapid freezing solution (UFS) for cryopreservation of day 2 mouse embryo. Results: We tested toxicity by exposing embryos to vitirification solutions and a ultrarapid freezing solution. The survival rates are 100%, 97.8%, 95.6% and 100% (EFS, VS14, DPS and UFS). After cultured for 96 hours, hatching rates of each group are 93.5% (no freezing), 95.6% (EFS), 86.4% (VS14), 93.0% (DPS), and 93.0% (UFS). There is no significant differences among groups. The survival rates after thawing cryopreserved embryos are 80.2%, 91.7%, 69.5%, 0% and 91.8% (slow freezing, EFS, VS14, DPS and UFS). Also cultured for 96 hours, the hatching rates are 93.5% (no freezing), 84.1% (slow freezing), 93.9% (EFS), 48.5% (VS14) and 70.1% (UFS). Conclusion: The survival rates of vitrification in EFS solution and ultrarapid freezing are higher than slow freezing (p<0.05). The hatching rate of vitrification in EFS solution cultured for 96 hours is highest, so vitrification of day 2 mouse embryos in EFS solution considered as more effective for cryopreservation.