猪滋养层干细胞样细胞的分离培养

Isolation and Cultivation of Porcine Trophoblastic Stem-like Cell Lines

滋养层干细胞(TSc)是形成胎盘组织细胞的前体细胞,与胚胎着床和胎盘形成密切相关。体外分离滋养层干细胞为研究滋养层的发育与功能提供了研究工具和基础。滋养层干细胞已经成功从小鼠附植前囊胚中获得,在猪上也有相关报道,但并没有关于6d囊胚中分离到猪滋养层干细胞(pTSc)的报道。本试验通过对6d孤雌囊胚透明带进行划口处理,饲养层和基础培养液中附加碱性成纤维生长因子(bFGF)和人白血病抑制因子(hLIF)分离猪滋养层干细胞样细胞。同时,通过采用形态学和基因表达分析的方法对获得的猪滋养层干细胞样细胞进行了初步鉴定。结果显示,本试验分离得到的细胞呈现上皮样细胞形态,细胞间结合紧密,边缘光滑,核质比较大,RT-PCR结果显示表达滋养层干细胞标记基因Cdx2,体现了滋养层干细胞特征。结果表明,本试验成功在6d孤雌囊胚分离得到猪滋养层干细胞样细胞,为后续研究猪滋养层发育提供了试验基础。

Trophoblast stem cells（TSc） were the precursors of the differentiated cells of the placenta.It was associated with embryo implantation.Isolating trophoblast stem cells in vitro provided research foundation for trophoblastic development.TSc had been successfully derivated from preimplantation embryo,there were also many reasearches about porcine trophoblast stem cells（pTSc）,but no reports indicated derivating 6 d pTSc.After broken the zona pellucida,we fixed the blastocyst on feeder under sterile condition.The porcine trophoblatic stem-like cell was intially analysed by morphogenesis and RT-PCR.We found the porcine trophoblastic stem-like cell combined tightly and present epithelioid-like cell.The clone was smooth,the ratio of the nucleus and cytoplasm was mass.The porcine trophoblastic-like cell was confirmed expressing Cdx2 gene by RT-PCR.We successfully derived the porcine trophoblastic stem-like cell from 6 d pathenogonidium blastcyst.The result provided some foundation for follow-up studies of porcine trophoblast development.