Journal of Chinese Physician 2011;13(8):1026-1029,1032
doi:10.3760/cma.j.issn.1008-1372.2011.08.005
Generation of human induced pluripotent stem cells from human skin fibroblasts
Zhixin JIANG ; Qinghua ZHANG ; Min AI ; Hang SHA ; Yi GAO ; Hai LU
Keywords
Skin/CY; Fibroblasts; Multipotent stem cells; Cell culture techniques
Country
China
Language
Chinese
Abstract
ObjectiveGeneration of human induced pluripotent stem cells from human skin fibroblasts.MethodsSox2, Klf4, Oct4, c-Myc were transfected into HSF cells with retrovirus, and then HSF cells was reprogrammed to iPS cells.Detecting cells endogenous and exogenous gene, analyzing karyotype,cells alkaline phosphatase staining and immunofluorescence staining, differentiating into teratomas in vivo and embryoid bodies in vitro were performed.ResultsiPS cell morphology was similar to embryonic stem cells (ES).The expression of Nanog, Oct4, Rex1, Sox2 in iPS cells were higher than HSF cells, and Sox2, Klf4, Oct4, c-Myc were silenced for the iPS cells.Exogenous genes were inserted into the nucleus of iPS cells, which was confirmed by 1% agarose gel electrophoresis.iPS cell karyotype was normal, alkaline phosphatase activity increased, ES cell-specific protein expressed.iPS cells were differentiated into a teratoma in vivo and embryoid bodies in vitro.ConclusionsiPS cells were similar to ES cells, which have pluripotency.
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