The survival of rabbit morulae preserved in liq-uid nitrogen after rapid thawing

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  • 凍結保存家と桑実はいの生存性に及ぼす急速融解の影響
  • トウケツ ホゾン カト ソウジツハイ ノ セイゾンセイ ニ オヨボス キュウソ

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The present study was undertaken to examine the effects of rapid thawing on the viability of rabbit morulae stored in liquid nitrogen. Embryos at morula stage were recovered at 60 to 65 h post coitum from the superovulated rabbits. The embryos were enclosed in a 1 ml capacity of plastic straw with PBS and 1.5 M-DMSO, and slowly cooled (1 C/min) to various temperatures ranging between -20 and -70C before transfer to liquid nitrogen and stored for 1 to 30 days. At thawing, the samples were slowly (4C/min) or rapidly (>1000 C/min) warmed. The other procedures for freezing and thawing were the same as that reported previously16). The frozen-thawed embryos were cultured for 2 days to determine the number of blastocysts developing in vitro. In some experiments, the frozen-thawed embryos were transferred directly after rapid thawing to the oviducts and/or uteri of recipient rabbits induced pseudopregnancy by the injection of HCG 2 and 4 h before (+) or 3 and 12 h after (-) the time of HCG injection in donor rabbits. We concluded that<BR>1. The incidence of rupture of zona pellucida in the rapidly thawed embryos was significantly (P <0.001) higher than that obtained in slow thawing.<BR>2. A high rate of survival in vitro (80-91%) of rapidly thawed embryos was obtained using the embryos had been transferred to liquid nitrogen from -30 to -50C. On the contrary, survival rate in vitro of slowly thawed embryos was low (0-53%) except in those had been subjected to slow cool-ing until -60C (90-91%).<BR>3. The chance to successful pregnancy after transfer of rapidly thawed morulae (1/15) was clearly less than that reported previously by using slow thawing method (7/8)23).

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