title: Production of mouse fibroblast growth factor 4 in E. coli and its application for isolation and maintenance of mouse trophoblast stem cells in vitro.
abstract: 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.
authors: Laboratory for Advanced Animal Cell Technology, Graduate School of Bioresource Sciences, Akita Prefectural University, Akita 010-0195, Japan.
authors: Yusuke
authors: Y
authors: Hosoi
authors: Yumi
authors: Y
authors: Ando
authors: Megumi
authors: M
authors: Arakawa
authors: Kano
authors: K
authors: Kasuga
authors: Ikuo
authors: I
authors: Kojima
authors: Masayuki
authors: M
authors: Kobayashi
mesh_terms: D000818:Animals; D019070:Cell Lineage / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005260:Female; D051519:Fibroblast Growth Factor 4 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D011247:Pregnancy; D061251:Primary Cell Culture / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
chemical_list: C496445:FGF4 protein, human; D051519:Fibroblast Growth Factor 4; D011994:Recombinant Proteins