생쥐 수정란의 핵이식후 체외발달에 관한 연구

Study on In Vitro Development of Mouse Embryos

Single nuclei from two-, four- and eight-cell mouse embryos were transplanted into enucleated two-cell mouse embryos by micromanipulation and sendai virum mediated fusion. no significant difference in successful injectin rate and fusion rate was found between the cell stages of nuclear donor embryos. There nuclear transplant embryos receiving different cell stage nuclei were cultured in vitro for 96 hours. 75.3% of 255 embryos receiving 2-cell nuclei, 68.2% of 236 embryos reciving 4-cell nuceli and 46.9% of 228 embryos receiving 8-cell nuclei were developed to blastocyst, respectively. The number of blastomeres was significantly(P<0.05) reduced in the embryos receiving 8-cell nuclei, compared with the embryos receiving 2-cell, 4-cell nuclei or the intact embryos. Also the size of blastocysts was significantly(P<0.05) smaller in the embryos receiving 8-cell nuclei, compared with the intact or other nuclear transplant embryos. These results suggest that single nuclei introduced into the enucleated two-cell embryos are able to support the in vitro development of the reconstituted embryos to blastocysts. The prominant retardation of blastocoele formation and cell division was shown in nuclear transplant embryos receiving eight-cell nuclei when they were cultured in vitro.