Preliminary Study On Differentiation Of Human Induced Pluripotent Stem Cells Into Epidermal-like Stem Cells In Vitro

Objective:This study focus on the culture and characteristics of human induced pluripotent stem cells and investigated the possibility for inducing the differentiation of induced pluripotent stem cell into the epidermal-like stem cells in vitro, so as to lay an experimental basis for provide new seed cells source of skin tissue engineering.Method:The induced pluripotent stem cells were plated on murine embryonic fibroblast feeder cells and expanded in human embryonic stem cells media contained basic fibroblast growth factor. The cells were passaged by collagenaseⅣdigestion method and observed under invert microscope. The expression of alkaline phosphatase was detected Based on the knowledge of human skin development, we pasted on human amniotic membrane with epithelial surface upward and covered the bottom of culture plate. Then the induced pluripotent stem cell were seeded on the amniotic membrane and cultured in human embryonic stem cells media without basic fibroblast growth factor. No amniotic membrane were used in control group.The expression of integrinβ1 and CK19 was detected by immunohistochemistry.Results:The cultured induced pluripotent stem cells grew well and stability with similar characteristics of human embryonic stem cells. These cells also expressed alkaline phosphatase. They formed embryoid body in feeder-free and suspension culture conditions. After 4 days of co-culture, the stem cell clones were formed on the surface of amniotic membrane, and part of the cells were integrinβ1 and CK19 positive. Most of the cells in control group died, and no integrinβ1 and CK19 positive cells were found. Conclusion:The cultured induced pluripotent stem cells grew well and stability with similar characteristics of human embryonic stem cells. The human induced pluripotent stem cells can differentiate into the epidermal-like stem cells by amniotic membrane in vitro. These results provide an experimental basis for improvement of induction study and further application to generate patient-specific induced pluripotent stem cells.