摘要
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。
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 serumfree 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 serum-free media.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2007年第2期1-5,10,共6页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家"863"计划项目(2005AA219050)
教育部重大项目(03160)
关键词
山羊
血清替代品
内细胞团
胚胎干细胞
免疫外科
goat serum replacement inner cell mass embryonic stem cell
immunosurgery
