【作者】 施巧婷； 王旭光； 王力虎； 刘贤侠； 杨梅； 郭志勤；

【Author】 SHI Qiao-ting, WANG Xu-guang, WANG Li-hu, LIU Xian-xia, YANG Mei, GHI Zhi-qin( Major Development Lab. of Veterinary Medical and Biological Technology, Xinjiang Academy of Animal Science, Urumqi, 830000; College of Animal Science, Xinjiang Agriculture University, Urumqi 830052; 3. Biology Department, Beying Argriculture College, Beijing 100206; 4. College of Animal Science, Shihezi University, Shihezi, 832000)

【机构】 新疆农业大学动物科技学院； 北京农学院生物系； 石河子大学动物科技学院； 农业部家畜繁育生物技术重点开放实验室；

【摘要】 血清饥饿和完全汇合对山羊皮肤成纤维细胞周期是否有影响?由流式细胞仪测定血清饥饿的细胞周期分布为G1/G0,S和G2+M期分别为79.9,7.5,11.6%,汇合细胞的周期分布分别为84.9,4.5,9.4%。本实验探讨了山羊新鲜卵丘细胞和培养冷冻后解冻的颗粒细胞作为核供体对核移植效率的影响,比较了颗粒细胞与成纤维细胞在饥饿和完全汇合时作为供体细胞对核移植效率的影响,以及血清饥饿与完全汇合培养的成纤维细胞的细胞周期,同时对比了电脉冲参数对核移植胚融合和发育的影响。结果表明新鲜卵丘细胞重构胚的融合率与培养颗粒细胞组差异显著,但卵裂率及桑囊胚率差异不显著。颗粒细胞作为核供体融合率和卵裂率均显著高于成纤维细胞,但桑囊胚率差异不显著。汇合培养与饥饿培养的细胞作为核供体在融合率、卵裂率、桑囊胚率差异均不显著。山羊重构胚电融合时电脉冲参数以电场强度1.2-1.6kv/cm,脉冲持续15μs与20μs,脉冲两次最适合重构胚的发育。更多还原

【Abstract】 The effect of serum starvation and confluent treatment on the cell cycle of goat skin ? Analysis of cell cycle distribution by fluorescence-activated cell sorting (FACS) showed that 79.9,7.5, 11.6% of serum stravation cycling cells were at Gl/GO , S and G2+M phase, respectively, and confluent cells for 84.9,4.5,9.4% reapectively. In this study we also tested the effects of the fresh and culture-frozen-thawed granulosa cells on nuclear transfer of goat. And compared the effects of serum stavation with conflunt cells of grnulosa and fibroblast cells on nuclear transfer of goat. And compared different electric pulse parameter on fuse and development of embryos of nuclear transfer. The results: the rate of fusion of fresh cells was significantly higher than that of culture-frozen-thawed cells(p<0.05), but cleavage rate and murulae/blastocyst rate had no significant difference. Fibroblast cells had significantly lower than granulosa cells in fusion and cleavage rate(p<0.05), but the murulae/blastocyst rate had no significant difference. The confluent and serum starvation cells as donor, the embryo had no significant difference in fusion, cleavage and murulae/blastocyst rate. And the reconstructed embryos were tested with 1.2-1.6kv/cm, 15-20 \is and 2 pulses electric activation had high activation rate.更多还原