喉上皮细胞条件液诱导胚胎干细胞分化为上皮细胞的实验研究

目的:探讨体外用喉上皮细胞条件培养液诱导胚胎干细胞分化为上皮细胞的方法及诱导条件。方法:将E14小鼠胚胎干细胞体外培养形成拟胚体后，在人喉上皮细胞条件培养液及因子诱导条件下共培养 14 d，形态学观察并采用免疫细胞化学方法检测上皮细胞的角蛋白Ck 4、14、19蛋白质水平的表达情况。结果:在诱导14 d的培养系统中较对照细胞可见更多上皮样细胞出现，免疫细胞化学染色显示上皮样细胞胞浆中可见角蛋白Ck 4、14、19均有表达。结论:胚胎干细胞在喉上皮细胞条件培养液诱导条件下可形成上皮样细胞，为组织工程制作人工喉上皮材料奠定了实验基础。

Directed differentiation of embryonic stem cells into epithelial cells by co-culture with human laryngeal epithelial cell conditioned medium

AIM: To explore the culture and inductive condition for the directed differentiation of embryonic stem cells into epithelial cells in human laryngeal epithelial cell conditioned medium.METHODS: E14 murine stem cells were induced into embryoid body in vitro first,then the cells of embryoid body were cultured in the induction system containing human laryngeal epithelial conditioned medium and epithelial cell growth factors.On day 14 of culture,samples were detected for the cytokeratins 4,14,19 expressions at protein level.Morphology of the cells was also observed under microscope.RESULTS: During the process of epithelial induction,cells with epithelial cell morphology were seen under contrast microscope.Immunofluorescent assay showed that cytokeratins 4,14,and 19 were all expressed at protein level in differentiated cell.CONCLUSION: This study suggests that epithelial cells could be induced from the directed differentiation of embryonic stem cells in the conditioned medium of human laryngeal epithelial cell cultures.This work has provided basic data for artificial laryngeal epithelia preparation.