Study on in vitro Fertilization, Embryonic Culture and Embryonic Rapid-Freezing in Mice
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Q813.7

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    Abstract:

    Objective\ To collect age\|specific embryos, increase the resources of embryos, and establish the bank of frozen embryos in mice. Methods\ The in vivo development and movement of zygotes, the in vitro maturation and fertilization of oocytes , the in vitro culture and rapid\|freezing of embryos were studied. Results\ (1) Zygotes developing to pronuclei occurred at 12-20h after injection of hCG;2\|cell, 4\|cell, and 8\|cell embryos formed at 42-48 h, 48-60 h, and 60-68 h, respectively. The embryos of the above stages were in the oviducts. Morula, early blastocysts, and blastocysts, which were in the uterus horns, formed at 75-70 h, 90-92 h, and 92-96 h, respectively. (2)Adding FSH and hCG to the culture medium could increase significantly the percentage of in vito fertilization of oocytes. Adding FCS and FSH and hCG simultaneously to the culture medium was better than adding hormone alone. FCS could increase significantly the rate of the embryo development. (3)Adding EDTA to the culture medium could effectively overcome the 2\|cell block in mice. The rates of embryos developed to 2\|cell stage and to 8\|cell stage were 100% and over 55%, respectively. Adding sodium lactate and pyruvate sodium to the culture medium could stimulate significantly the embryonic development. (4) After frozen in the freezing solution (D\|PBS+glycerol+sucrose), and thawed in DPBS +sucrose solution, the survival rate of morula and early blastocysts were 69.3% and 60.4%, respectively. Conclusion\ The present results laid the preliminary basis for the establishment of embryo bank in mice.

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  • Received:
  • Revised:June 12,2001
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