Abstract Research of the induced pluripotent stem cells(iPSCs) by inducing the SIM-6-thiogunanie-oualiain(STO) cells. Lentivirus vector FUW-OSKM which included octamer-binding transcription factor 4(Oct4), SRY (sex determining region Y)-box 2(Sox2), Kruppel-like factor 4 (Klf4) and c-Myc transfected to 293FT cells by calcium phosphate method. Then, using the virus infected STO cells after lentivirus was packaged and the cells were cultured in the ESC media. The clones were isolated on the fifteenth day cultured on feed layer and none feed layer culture dishs.Through the biological characteristics analysis, we found the cells named STO-iPSCs had the typical morphological characteristics of embryonic stem cells, had a positive result by alkaline phosphatase staining,expressed embryonic stem cells marker Nanog and Oct4 by the immunocytochemistry and high quantitys of Nanog and Oct4 mRNAs by qPCR, especially could differentiate to nerve cells in vitro. In conclution, we obtained STO-iPSCs and stablished the STO-iPSCs training system without raising layer.
|
Received: 12 March 2012
|
|
|
|
|