Generation of human induced pluripotent stem cells from human skin fibroblasts / 中国医师杂志

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.