摘要
以CEF细胞、MEF细胞和STO细胞为饲养层,用BRL-3A条件培养基培养寿光鸡Ⅹ期胚盘细胞,并对获得的细胞克隆进行了鉴定,证明其具有ES细胞的克隆形态、PAS染色阳性、AKP染色阳性、能分化为多种类型的细胞、能参与受体胚胎的发育并形成羽色嵌合体。实验结果表明:BRL-3A条件培养基能促进鸡ES细胞克隆的形成,用饲养层比不用饲养层更有利于克隆的形成,且STO细胞和MEF细胞饲养层的培养效果要好于CEF细胞饲养层(P<0.05);挑克隆传代法比全消化法更有利于克隆的形成和未分化状态的维持(P<0.05)。
The culture system of chicken embryonic stem （ES） cells by buffalo rat liver cells conditioned medium （BRL-CM） was optimized. Stage X blastoderm cells of Shouguang chicken were cocultured with feeder layers （CEF, MEF and STO） in BRL-CM. The clone was identified by morphology, PAS staining, AKP staining, constructing chimeras and differentiation in vitro. Results showed that BRL-CM could stimulate chicken ES cells proliferation. Feeder layers could maintain the proliferation and undifferentiation of chicken ES colonies. Compared with CEF feeder layers, MEF and STO feeder layers were more ideal in culture of chicken ES cells （P〈0.05）. The passage method of picking colonies was more efficient than trypsin digestion （P〈0.05）.
出处
《细胞生物学杂志》
CSCD
2008年第4期532-536,共5页
Chinese Journal of Cell Biology
基金
北京市自然科学基金重点资助项目(No05E143)~~
