体外诱导胚胎干细胞来源表皮干细胞向汗腺细胞分化的研究

目的:探讨体外诱导胚胎干细胞来源表皮干细胞向汗腺上皮分化可行性及条件,为汗腺组织工程探索新的种子细胞提供来源。方法:体外分离培养、扩增并鉴定人汗腺细胞。将胚胎干细胞培养于人羊膜表面诱导其向表皮干细胞转化后,与人汗腺细胞直接共同培养,诱导其分化为汗腺细胞。采用倒置显微镜进行形态学观察,并用免疫细胞化学染色方法检测诱导分化结果。结果:体外培养的人汗腺细胞CK19、CEA阳性表达。培养于人羊膜表面的胚胎干细胞β1整合素呈强阳性表达,符合表皮干细胞的表面抗原标志。经与人汗腺细胞共培养2周后,有部分胚胎干细胞来源的表皮干细胞表达CEA,表明通过汗腺细胞分泌的细胞因子、细胞间直接接触等可能的作用途径,使其表型向汗腺细胞表型转化。结论:胚胎干细胞来源表皮干细胞可能成为汗腺组织工程种子细胞的重要来源。

Study on the differentiation of embryonic stem cells derived epidermal stem cells into sweat gland cells in vitro

Objective:To investigate the feasibility and condition of embryonic stem cells(ESCs) derived epidermal stem cells differentiate into sweat gland cells(SGCs),provide new source of seed cells for sweat gland engineering.Methods:SGCs were isolated,expended in vitro,and identified in advance.Epidermal stem cells were harvested from human amnion epithelial surface where ESCs were seeded and induced,and were induced differentiate toward SGCs by co-culture with them.Morphological observation was performed through inverted microscope every day.The inductivity was determined by immunocytochemistry.Results:The cultured SGCs expressed CK19 and CEA.After cultured on human amnion epithelial surface,the ESCs extremely expressed integrin β1,a surface marker of epidermal stem cells.Part of the ESCs derived epidermal stem cells expressed CEA after co-culture with SGCs,suggesting a phenotype transformation which maybe caused by the effect of SGCs secreted cytokine or/and cell-to-cell contact.Conclusion:The ESCs derived epidermal stem cells may become an important new source of seed cells for sweat gland engineering.