无血清培养山羊内细胞团的研究

对32只关中奶山羊进行超排处理,获得胚胎263枚,其中囊胚122枚。采用机械法、酶消化法和免疫外科法分离囊胚83枚,得到内细胞团(ICM)65个。将ICM培养于小鼠胎儿成纤维细胞饲养层上,研究含血清和无血清条件下ICM的增殖规律,并比较添加不同生长因子对ICM增殖的影响。结果表明,无血清培养时,ICM细胞增殖相对较慢,分化减少;添加LIF、bFGF、两种同时添加或均不添加对原代ESC样集落形成率的影响不显著(P>0.05),但添加10 ng/mL bFGF稍微有助于ICM增殖。山羊ICM适宜于无血清培养。

The study of serum-free culture of goat inner cell mass

A total of 32 Guanzhong dairy goats were superovulated with a result of 263 embryos including 122 blastocysts,65 intact inner cell mass(ICM) obtained from 83 blastocysts by mechanical incision,enzymatic digestion,or immunosurgery methods.The propagation characteristics of ICM were studied both in serum-containing and serum-replacement media on a feeder layer of mitomycin-inactivated mouse embryo fibroblasts(MEF).The result indicates that the differentiation of ICM decreased in serum-free media,although the proliferation of ICM was slower in the condition of serum-free than in the serum-medium.The proliferation condition of ICM(total 30) were cultured by addition of different growth factors in serum-free media,there were no statistical differences of primary colony formation among the four groups(added to LIF,bFGF,both or none) on MEF cell feeder layer(P>0.05),however,addition of 10 ng/mL bFGF may slightly promote the proliferation of goat ICM.These results suggest that it is suitable to culture goat ICM in serumfree media.