Abstract: Objective To optimize the method for establishment of induced pluripotent stem cells from human foreskin fibroblasts. Methods We used GFP as an indicator to determine the optimized dose of the retrovirus during the reprogramming of human foreskin fibroblasts into human induced pluripotent stem cells (iPSCs) by Yamanaka’s classical strategy with the combination of the small-molecule compounds VPA, a histone deacetylase inhibitor. The characteristics of established iPSCs were tested via alkaline phophatase (AP) staining, immunofluorescent detection of human embryonic stem (ES) cells surface antigens, and RT-PCR analysis of three germ layers specific gene expression in cell during differentiation in vitro. Results Using GFP expression as an indicator of the retrovirus transduction, we established a high efficient system to successfully reprogramme human foreskin fibroblasts into induced pluripotent stem cells. The iPSCs showed positive for alkaline phosphatase, expressed high levels of human ES cells pluripotency markers such as OCT4,TRA-1-60 and TRA-1-81. RT-PCR analysis indicated that the germ layers differentiation related gene expression was detected in the embryonic body when derived from these hiPS cells. BR>Conclusion Our results showed that we successfully generated hiPS cells from adult foreskin fibroblasts in the indication of GFP with the combination of the small-molecule compoun

Key words: Stem cell, Differentiation, Foreskin fibroblast, Green fluorescent protein, Immunofluorescence, Human