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Biomaterials ; 35(24): 6259-67, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24811263

ABSTRACT

Nanofibrous gelatin substrates are suited for long-term expansion of human pluripotent stem cells (hPSCs) under feeder- and serum-free culture conditions. A combinatorial library with different sets of processing parameters was established to assess the culture performance of hPSCs on nanofibrous substrates in terms of cell adhesion and growth rate, using Matrigel as control. Then, the optimal conditions were applied to long-term expansion of hPSCs with several cell lines, showing a maintained pluripotency over more than 20 passages without introducing any abnormal chromosome. In addition, this approach allowed us to avoid enzymatic disassociation and mechanic cutting during passages, thereby promoting a better hPSC culture and long-term expansion.


Subject(s)
Cell Culture Techniques/methods , Gelatin/pharmacology , Nanofibers/chemistry , Pluripotent Stem Cells/cytology , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Cell Shape/drug effects , Cells, Cultured , Embryonic Stem Cells/cytology , Embryonic Stem Cells/drug effects , Embryonic Stem Cells/ultrastructure , Gene Expression Regulation/drug effects , Humans , Integrins/metabolism , Karyotyping , Pluripotent Stem Cells/drug effects , Pluripotent Stem Cells/ultrastructure , RNA, Messenger/genetics , RNA, Messenger/metabolism
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