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Cloning and expression of mLIF and the effects of mLIF on maintaining chicken primordial germ cells in the undifferentiated state

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Author(s): YANG Na-Na | ZHU Jing | SHI Wei-Feng | MENG Chun-Hua | DU Li-Xin

Journal: Acta Zoologica Sinica
ISSN 0001-7302

Volume: 54;
Issue: 5;
Start page: 903;
Date: 2008;

ABSTRACT
In order to establish the best cultivation system, we constructed eukaryotic expression vector pSecTag-mlif(sp-), transfected chicken PGCs(Primordial germ cells) and CEF(Chicken embryonic fibroblast) cells with pSecTag-mlif(sp-) by the method of lipofectamine mediation, and collected the supernatant after centrifugating the cellular medium. We surveyed mLIF(Mice leukemia inhibitory factor) expression in cellular medium by means of of western-blotting. We cultured PGCs using seven different methods including three control groups, which all utilized CEF cells as feeder. The PGC clones cultured in the forth group and the fifth group proliferated perfectly. The second passage CEG clones cultured in the third group proliferated very well at first, but the CEG clones began differentiating three days later. We sucsesfully transfected chicken PGCs and CEF cells with pSecTag-mlif(sp-), and the mLIF expressed by pSecTag-mlif(sp-) can maintain PGCs cells in an umdifferentiated state [Acta Zoologica Sinica 54(5): 903 – 908, 2008].
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