Generation of human induced pluripotent stem cells from umbilical cord matrix cells

Abstract

Objective To generate of human induced pluripotent stem cells from umbilical cord matrix cells(UMC).Methods Sox2 and Klf4 and Oct4 and c-Myc were transfected into UMC cells with retrovirus,and thcn UMC cells was reprogrammed to iPS cells.Gene expression was confirmed with RT -PCR and the integration was confirmed with cell karyotype.iPS cells were further validatcd with cell alkaline phosphatase detection and immunofluorescence staining,differentiating into teratomas in vivo and embryoid bodies in vitro.Results iPS cells were similar to embryonic stem cells (ES).The expression of Nanog,Oct4,Rex1 and Sox2 in iPS cells were higher than UMC cells,and Sox2,Klf4,Oct4,c-Myc silenced in iPS cells.Exogenous genes were inserted into the nucleus of iPS cells,which was confirmed by 1％ agarose gel electrophoresis.iPS ccll karyotype was normal,alkaline phosphatase activity increased,ES cell-specific proteins,including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog,were expressed.iPS cells were differentiated into a teratoma in vivo and embryoid bodies in vitro.Conclusions iPS cells were similar to ES cells,which had pluripotency. Key words: Umbilical cord/CY; Stromal cells/CY; Multipotent stem cells