【作者】 桑润滋； 韩建永； 孙国杰； 金东航； 王志刚； 李俊杰；

【Author】 SANG Run-zi, HAN Jian-yong, SUN Guo-jie, JIN Dong-hang, WANG Zhi-gang, LI Jun-jie(College of Animal Science and Technology, Agricultural University of Hebei, Bonding 071001China)

【机构】 河北农业大学动物科技学院；

【摘要】 本试验就胚龄、饲养层、生长因子及培养系统等因素对山羊胚胎干细胞分离克隆效率的影响进行研究,结果表明,在2种不同饲养层上,囊胚的脱带时间均短于桑椹胚,囊胚的脱带率均高于桑椹胚,而饲养层的种类对胚胎的脱带时间以及脱带率影响不大。脱带切割囊胚不论在PMEF还是在PGEF饲养层上,其贴壁时间均短于脱带整胚及孵化囊胚,而贴壁率高于脱带整胚,与孵化囊胚相似。脱带整胚及脱带切割胚在PMEF饲养层上所获类ES细胞只能维持3代,而在PGEF饲养层上,脱带切割半胚和孵化囊胚所获类ES细胞传至5代。对山羊45d胎儿原始生殖细胞(PGCs)分离克隆结果表明,以PMEF、PGEF饲养层以及与生殖嵴周围组织共同培养的方法,添加细胞因子,均能获得类EG集落。更多还原

【Abstract】 The effect of embryonic age, feeder layer, growth factor, cultural system on the efficiency of isolation and cloning embryonic stem cell of goat was studied. Results showed that the dissociation time of blastocysts from zone pcllucida was shorter than that of morulae, and the dissociation rate was higher, whenever on the feeder layer PMEF or on PGEF. There was no significant effect of feeder layer on the time and rate of dissociation. The attachment time of bisected dissociated blastocyst with ICM was shorter than that of intact dissociated blastocysts and hatched blastocysts, and the attachment rate of bisected embryos was was as high as that of hatched blastocysts, but higher than intact blastocysts. The third passage of ES-like cells were obtained from the intact blastocysts and bisected embryos on the PMEF feeder and the fifth passage of ES-like cells were obtained on the PGEF feeder. Isolation and cloning of primordial germ cells from day 45 fetuses of goat indicated that EG-like cells could be derived both from culture on PMEF, PGEF feeder layer and co-culture in medium with cell growth factors.更多还原