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dc.contributor.authorLu, Huai-Enen_US
dc.contributor.authorTsai, Ming-Songen_US
dc.contributor.authorYang, Yao-Chenen_US
dc.contributor.authorYuan, Chen-Chingen_US
dc.contributor.authorWang, Tzu-Haoen_US
dc.contributor.authorLin, Xi-Zhangen_US
dc.contributor.authorTseng, Ching-Pingen_US
dc.contributor.authorHwang, Shiaw-Minen_US
dc.description.abstractGeneration of induced pluripotent stem (iPS) cells from somatic cells has been successfully achieved by ectopic expression of four transcription factors, Oct4, Sox2, Klf4 and c-Myc, also known as the Yamanaka factors. In practice, initial iPS colonies are picked based on their embryonic stem (ES) cell-like morphology, but often may go on to fail subsequent assays, such as the alkaline phosphate (AP) assay. In this study, we co-expressed through lenti-viral delivery the Yamanaka factors in amniotic fluid-derived (AF) cells. ES-like colonies were picked onto a traditional feeder layer and a high percentage AF-iPS with partial to no AP activity was found. Interestingly, we obtained an overwhelming majority of fully stained AP positive (AP+) AF-iPS colonies when colonies were first seeded on a feeder-free culture system, and then transferred to a feeder layer for expansion. Furthermore, colonies with no AP activity were not detected. This screening step decreased the variation seen between morphology and AP assay. We observed the AF-iPS colonies grown on the feeder layer with 28%AP+ colonies, 45% AP partially positive (AP+/-) colonies and 27% AP negative (AP-) colonies, while colonies screened by the feeder-free system were 84%AP+ colonies, 16% AP+/- colonies and no AP- colonies. The feeder-free screened AP+ AF-iPS colonies were also positive for pluripotent markers, OCT4, SOX2, NANOG,TRA-1-60,TRA-1-81,SSEA-3 and SSEA-4 as well as having differentiation abilities into three germ layers in vitro and in vivo. In this study, we report a simplistic, one-step method for selection of AP+ AF-iPS cells via feeder-free screening. (C) 2011 Elsevier Inc. All rights reserved.en_US
dc.subjectAmniotic fluiden_US
dc.subjectInduced pluripotent stem cellsen_US
dc.subjectFeeder freeen_US
dc.subjectAlkaline phosphataseen_US
dc.titleSelection of alkaline phosphatase-positive induced pluripotent stem cells from human amniotic fluid-derived cells by feeder-free systemen_US
dc.identifier.journalEXPERIMENTAL CELL RESEARCHen_US
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
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