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Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

Article DOI: 10.3791/3416-v • 13:24 min • February 20th, 2012
February 20th, 2012 •
1,2, 1, 3, 3, 1, 1
1Department of Molecular Carcinogenesis and Center for Cancer Epigenetics, University of Texas M.D. Anderson Cancer Center, 2Department of Cell Biology, Poznan University of Medical Sciences, 3Department of Molecular Biology, The Scripps Research Institute

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We present a protocol for efficient reprogramming of human somatic cells into human induced pluripotent stem cells (hiPSC) using retroviral vectors encoding Oct3/4, Sox2, Klf4 and c-myc (OSKM) and identification of correctly reprogrammed hiPSC by live staining with Tra-1-81 antibody.

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Selecting Isolating Colonies Human Induced Pluripotent Stem Cells Reprogramming Adult Fibroblasts Retroviral Vectors Oct3/4 Sox2 Klf4 C-myc Sodium Butyrate Efficient Transduction Protocol Virus-containing Media Live Immunostaining Tra-1-81 Pluripotent Cells Manually Passaged Matrigel Plates Feeder-free Conditions HES-like Morphology Expansion Cell Lines Pluripotency Markers TRA-1-60 SSEA-4 Oct3/4 Sox2 Nanog
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