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This study was undertaken to determine the survival of rabbit fertilized eggs after long term storage in liquid nitrogen. Eggs at the 8-16 cell stage were recovered at 45 h post coitum from mature Japanese White rabbits which had been induced to superovulate by treatment with FSH and HCG. The eggs were transferred into 0.1 ml PB1 supplemented with 50% rabbit serum contained in a 1 ml capacity plastic straw. The eggs were equilibrated with 1.5 M DMSO at 37°C for 15 min. The samples were cooled to -76°C at 1°C/min and transferred to liquid nitrogen vapor at -100°C for 5 min. The samples were finally transferred to liquid nitrogen at -196°C and preserved for 210, 4964 and 360463 days, respectively. The samples were warmed from -70°C to -10°C at approximately 4°C/min. At thawing, the DMSO in the medium was diluted in a step-wise manner with freezing medium at 37°C. The eggs were washed with Ringer's solution containing 20% rabbit serum and cultured for 45 days to determine the number of blastocysts developing <I>in vitro</I>. In the other experiments, the frozen-thawed eggs were transferred directly to the oviducts of synchronous pseudopregnant does which allowed to go to term. Statistical significance was determined by the X<SUP>2</SUP> test.<BR>1. The proportions of eggs which appeared morphologically normal after thawing of eggs preserved for 4964 and 360463 days were 82.5 and 71.8%. These figures were not signficantly different from that obtained after thawing of eggs preserved for 210 days, 82.1%. 2. The proportions of eggs developed into blastocysts after culture of frozen-thawed eggs preserved for 4964 and 360463 days were 77.8 and 62.5%. These were not significantly different from that obtained after culture of eggs preserved for 210 days, 63.4%. 3. The proportions of eggs developed into live young after transfer of eggs preserved for 4964 days and 360463 days, 18.5 and 17.9%, were comparable to that obtained after transfer of eggs preserved for 210 days, 16.7%.
- 日本繁殖生物学会の論文
日本繁殖生物学会 | 論文
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