供体核种类对水牛核移植效果的影响

采用电融合法和直接注射法进行核移植,以比较不同来源和不同类型水牛供体细胞的核移植效果。当用电融合法进行核移植时,成体耳部成纤维细胞的融合率(46.0%)、卵裂率(53.9%)和囊胚发育率(10.9%),均显著低于胎儿成纤维细胞(64.5%,70.1%和21.6%)、胎儿皮肤细胞(71.5%,70.8%和22.1%)以及卵巢颗粒细胞(88.2%,79.1%和25.5%);后三种细胞间的卵裂率和囊胚发育率无显著差异,但卵巢颗粒细胞的融合率显著高于胎儿成纤维细胞和胎儿皮肤细胞(P<0.05)。当用直接注射法进行核移植时,胎儿皮肤细胞注核后的存活率(78.2%)、卵裂率(41.6%)均显著低于成体耳部成纤维细胞(88.6%和57.4%)和胎儿成纤维细胞(85.8%和65.0%)。胎儿成纤维细胞注核后的卵裂率和囊胚发育率(23.3%)均显著高于胎儿皮肤细胞(10.1%)和成体耳部成纤维细胞(12.0%)。结果表明:(1)采用直接注核法进行核移植时,胎儿成纤维细胞作为核供体较为合适,而采用电融合法进行核移植时,用颗粒细胞作核供体更为适宜;(2)胎儿成纤维细胞的核移植效果好于成体成纤维细胞。

Effects of donor cell categories on the development of buffalo embryos derived from nuclear transfer

The development of nuclear transfer(NT)embryos derived from adult ear fibroblasts, fetal fibroblasts, fetal skin cells and ovary granulosa cells were compared. When electric fusion method was used for nuclear transfer, the fusion rate (46.0%), cleavage rate (53.9%) and blastocyte development rate (10.9%) of adult ear fibroblasts were significantly lower than those of fetal fibroblasts (64.5%, 70.1%, 21.6% respectively), fetal skin cells (71.5%, 70.8%, 22.1% respectively) and ovary granulosa cells (88.2%, 79.1%, 25.5% respectively). There was no significant difference in the cleavage and blastocyst development rate of reconstituted embryos among the later three donor cells. However, the fusion rate of ovary granulosa cells was significantly higher than that of the fetal fibroblasts and fetal skin cells (88.2% vs 64.4%, 71.5%,P<0.05). When direct injection was used for nuclear transfer, the survive rate (78.2%) and cleavage rate (41.6 %) of fetal skin cells were significantly lower than those of adult ear fibroblasts (88.6%, 57.4%) and fetal fibroblasts (85.8%, 65.0%). The cleavage rate and blastocyte development rate (23.3%) of fetal fibroblasts were significantly higher than that of fetal skin cells (10.1%) and adult ear fibroblasts (12.0%). These results show that granulosa cells are suitable for nuclear transfer by electric fusion, however, the fetal fibroblasts are suitable for nuclear transfer by direct injection.