RRC ID 67425
著者 Zhou Y, Mao H, Joddar B, Umeki N, Sako Y, Wada K, Nishioka C, Takahashi E, Wang Y, Ito Y.
タイトル The significance of membrane fluidity of feeder cell-derived substrates for maintenance of iPS cell stemness.
ジャーナル Sci Rep
Abstract The biological activity of cell-derived substrates to maintain undifferentiated murine-induced pluripotent stem (iPS) cells was correlated to membrane fluidity as a new parameter of cell culture substrates. Murine embryonic fibroblasts (MEFs) were employed as feeder cells and their membrane fluidity was tuned by chemical fixation using formaldehyde (FA). Membrane fluidity was evaluated by real-time single-molecule observations of green fluorescent protein-labeled epidermal growth factor receptors on chemically fixed MEFs. Biological activity was monitored by colony formation of iPS cells. Treatment with a low concentration of FA sustained the membrane fluidity and biological activity, which were comparable to those of mitomycin C-treated MEFs. The biological activity was further confirmed by sustained expression of alkaline phosphatase, SSEA-1, and other pluripotency markers in iPS cells after 3-5 days of culture on FA-fixed MEFs. Chemical fixation of feeder cells has several advantages such as providing ready-to-use culture substrates without contamination by proliferating feeder cells. Therefore, our results provide an important basis for the development of chemically fixed culture substrates for pluripotent stem cell culture as an alternative to conventional treatment by mitomycin C or x-ray irradiation.
巻・号 5
ページ 11386
公開日 2015-6-12
DOI 10.1038/srep11386
PII srep11386
PMID 26065582
PMC PMC4464345
MeSH Animals Cell Culture Techniques / methods* Feeder Cells / cytology Feeder Cells / metabolism* Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / metabolism* Membrane Fluidity* Mice
IF 3.998
リソース情報
ヒト・動物細胞 iPS-MEF-Ng-20D-17(APS0001)