The significance of membrane fluidity of feeder cell-derived substrates for maintenance of iPS cell stemness.
The significance of membrane fluidity of feeder cell-derived substrates for maintenance of iPS cell stemness.
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DOI：
10.1038/srep11386
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发表时间：
2015-06-12
影响因子：
4.6
通讯作者：
Ito Y
中科院分区：
文献类型：
--
作者：
Zhou Y;Mao H;Joddar B;Umeki N;Sako Y;Wada K;Nishioka C;Takahashi E;Wang Y;Ito Y
细胞来源的底物维持未分化的小鼠多能干细胞(IPS)的生物活性与膜流动性作为细胞培养底物的新参数相关。以小鼠胚胎成纤维细胞(MEF)为饲养细胞，用甲醛(FA)化学固定法调节其膜流动性。通过实时单分子观察绿色荧光蛋白标记的表皮生长因子受体在化学固定的MEF上的膜流动性。IPS细胞集落形成法检测其生物学活性。低浓度FA处理维持了膜的流动性和生物活性，与丝裂霉素C处理的MEF相当。在FA固定的MEF上培养3-5天后，iPS细胞持续表达碱性磷酸酶、SSEA-1和其他多能标志物，进一步证实了其生物学活性。饲养层细胞的化学固定有几个优点，例如提供即用的培养底物，而不会受到增殖的饲养层细胞的污染。因此，我们的结果为开发化学固定的多能干细胞培养底物以替代传统的丝裂霉素C或X射线照射治疗提供了重要的基础。
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.
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DOI：
10.1126/science.1191035
发表时间：
2010-08-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gilbert PM;Havenstrite KL;Magnusson KE;Sacco A;Leonardi NA;Kraft P;Nguyen NK;Thrun S;Lutolf MP;Blau HM
通讯作者：
Blau HM
影响因子：
6.7
作者：
Lee, Jae Hee;Lee, Eun Ju;Lim, Jeong Mook
通讯作者：
Lim, Jeong Mook
DOI：
10.1186/2045-9769-2-7
发表时间：
2013-01-01
期刊：
Cell regeneration (London, England)
影响因子：
--
作者：
Lambshead, Jack W;Meagher, Laurence;Laslett, Andrew L
通讯作者：
Laslett, Andrew L
影响因子：
6
作者：
Lim, Mei Ling;Jungebluth, Philipp;Macchiarini, Paolo
通讯作者：
Macchiarini, Paolo
影响因子：
4.8
作者：
Carter, RE;Sorkin, A
通讯作者：
Sorkin, A