金鱼胚胎细胞超低温保存及其发育能力的研究

CRYOGENIC PRESERVATION OF BLASTOMERES OF GOLD FISH (CA.RASSIUS AURATUS) AND ITS DEVELOPMENTAL POTENTIAL

将金鱼囊胚期的细胞分离后,保存于-196℃液氮中,并对保存后化冻的细胞用细胞核移植的方法检测了其细胞核发育的能力。本文首次证明了经冷冻后的细胞核被移至去核卵中能发育成正常的个体。并证明在冷冻液中加入胶原蛋白可以提高化冻后细胞的存活率。本实验的结果为长期保存端黄卵的各种鱼类的基因库提供了新的途径。

Individual blastomeres of gold fish were isolated at blastula stage and preserved in liquid nitrogen at -196℃ with the additives of DMSO and collagen in the medium for cryogenic storage. The effects of 10% DMSO mixed with collagen at various concentration as a protective reagent on cell damage in the medium were tested. It indicated that the percentage of viable cell could be increased significantly at 1-2% concentration of collagen.The developmental potential of frozen blastomeres was tested by nuclear transplantation. Blastomeres after storage at -196℃ were thawed and transplated into enucleated unfertilized eggs of gold fish. Normal fish were obtained from them. These results suggest that this method will provide a new avenue for preservation of gene pool of teleosts and it is probably available for others which belong to telolecithal eggs as well.