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首页> 外文期刊>The Journal of Reproduction and Development >Production of Mouse Fibroblast Growth Factor 4 in E. coli and Its Application for Isolation and Maintenance of Mouse Trophoblast Stem Cells In Vitro
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Production of Mouse Fibroblast Growth Factor 4 in E. coli and Its Application for Isolation and Maintenance of Mouse Trophoblast Stem Cells In Vitro

机译：大肠杆菌中小鼠成纤维细胞生长因子4的生产及其在体外分离和维持小鼠滋养细胞干细胞中的应用

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Fibroblast growth factor 4 (FGF4) promotes isolation of trophoblast stem (TS) cells from mouse blastocysts and maintenance of TS cells in an undifferentiated state in vitro . To date, commercially available, bacterially expressed human FGF4 (RhFGF4) has been used generally for this purpose. In this study, HismFGF4, a 6x histidine-tagged mouse FGF4, was produced in E. coli and purified using heparin column chromatography. We demonstrated that HismFGF4 (25 ng/ml) more efficiently generates mouse TS cells from a single blastocyst than RhFGF4 (25 ng/ml) and that TS cells isolated and maintained with HismFGF4 retained their ability to differentiate into the trophoblast cell lineage in vitro . In addition, TS cells cultured with HismFGF4 (25 ng/ml) were maintained in an undifferentiated state better than with RhFGF4 (25 ng/ml). To the best of our knowledge, this is the first application of a mouse FGF4 derivative for isolation and maintenance of mouse TS cells.
机译：成纤维细胞生长因子4（FGF4）促进从小鼠胚泡中分离滋养层干细胞（TS），并在体外以未分化状态维持TS细胞。迄今为止，可商购的细菌表达的人FGF4（RhFGF4）已普遍用于此目的。在这项研究中，HismFGF4是6x组氨酸标签的小鼠FGF4，在大肠杆菌中生产，并使用肝素柱色谱法纯化。我们证明了HismFGF4（25 ng / ml）比RhFGF4（25 ng / ml）从单个胚泡更有效地产生小鼠TS细胞，并且用HismFGF4分离并维持的TS细胞在体外具有分化为滋养层细胞谱系的能力。另外，用HismFGF4（25 ng / ml）培养的TS细胞比用RhFGF4（25 ng / ml）更好地保持未分化状态。据我们所知，这是小鼠FGF4衍生物首次用于分离和维持小鼠TS细胞的应用。

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