用BRL-3A条件培养基培养鸡胚胎干细胞的初步研究  

Preliminary Study on Culture of Chicken Embryonic Stem Cells by Buffalo Rat Liver Cells Conditioned Medium

作　　者：孟春花[1] 张传生[2] 杨娜娜[3] 王子玉[4] 曹少先[1] 杜立新[5] 

机构地区：[1]江苏省农业科学院畜牧研究所,南京210014 [2]河北科技师范学院动物科学系,昌黎066600 [3]泰山医学院生命科学研究所,泰安271000 [4]南京农业大学动物科技学院,南京210095 [5]中国农业科学院北京畜牧兽医研究所,北京100094

出　　处：《细胞生物学杂志》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)~~

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）.

关 键 词：鸡 胚盘细胞 胚胎干细胞 大鼠肝细胞条件培养基 

分 类 号：Q813[生物学—生物工程] Q959.837