摘要
虽然ＥＳ细胞技术的应用十分广泛，对ＥＳ细胞多能性本质的研究还不是很深入，体外培养的ＥＳ细胞群体的不均一性加大了这方面研究的难度。报道了对ＥＳ细胞群体中未分化细胞的特异标记，ｏｃｔ－４是一个在小鼠胚胎所有全能性细胞和未分化的ＥＳ细胞中特异表达的基因，将报告基因βｇｅｏ插入ｏｃｔ－４基因转录元件中构建了标记载体ｐＧ１８ＮＧ，转染ＥＳ细胞ＭＥＳＰＵ２２和 ＭＥＳＰＵ１３后获得了稳定整合的细胞克隆。经体外培养、诱导分化、嵌合体制作等实验，证明利用该载体对ＥＳ细胞中的未分化细胞成功进行了标记，该标记在体内、体外都是有效的。
Despite the wide application of ES cell technology, little is known about the pluripotent nature of ES cells. This is partly due to the heterogeneity of ES cell population in culture. This report described the specific labelling of undifferentiated cells in ES cell lines. Oct-4 gene is specifically expressed in all the totipotent cells in mouse embtyos and undifferentiated ES cells. A constructed pG18NG was obtained by inserting the reporter geneβgeo into the oct-4 transcription elements. ES cell lines MESPU22 and MESPU13 were transfected with this construct and stable integrated cell clones were selected out. With the experiments of in vitro cultivation, differentiation and chimeras production, it was confirmed that we have successfully labelled the undifferentiated cells in ES cell lines, and this label was both valid in vitro and in vivo.
基金
国家自然科学基金资助(重点项目39730250)~~
