液体窒素(-196°C)に凍結されたマウスの8細胞卵および桑実胚の生存性に影響をおよぼす要因

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  • Factors Involved in the Survival of Mouse Eight-cell and Morula Embryos Frozen to -196°C
  • エキタイ チッソ 196ドC ニ トウケツサレタ マウス ノ 8 サイボウ ラ

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The present experiments have been disigned to examine the effects of cryoprotective agents, cooling rates, warming rates, age of female mice, and natural ovulation and super-ovulation on the survival of Inouse eight-cell and morula embryos frozen to -196°C. The methods used in these experiments were those reported previously by Whittingham et al.5) with some exceptions. The results were as follows. 1. No mouse embryos developed to the blastocysts stage after freezing and thawing in the presence of glucose, sucrose or polyvinylpyrrolidone. Whereas the embryos survived in the presence of glycerol or dimethyl sulfoxide (DMSO) and the highest survival was obtained in the medium of 1.2M DMSO (p<0.01).2. The highest survival was obtained with cooling rates of 0.2-0.5°C/min (p<0.01) and none survived after cooling rates of or 10°C/min. 3. The high survival was obtained with warming rates of 2 and 15°C/min and none survived after warming rate of 180°C/min.4. Similar proportions of embryos obtained from female mice aged 8 to 10 and 40 to 45 weeks survived after freezing and thawing. 5. Embryos obtained from naturally ovulated and superovulated mice showed the similar survival after freezing and thawing. 6. Similar proportions of eight-cell and morula embryos survived after freezing and thawing. 7. Forty blastocysts, obtained after freezing and thawing eight-cell eggs and culturing for a further 48 hours, were tranferred to the left uterine horns of five females on day 3 of pseudopregnancy. Two females were pregnant; the one female had three normal looking fetuses and the other female had four fetuses.

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