胚胎干细胞定向诱导为结膜上皮样细胞的实验研究

目的：研究体外诱导小鼠胚胎干细胞(embryonic stem cell,ESC)分化为结膜上皮细胞的可能性及条件，为构建组织工程化结膜提供新型的种子细胞奠定基础。方法：以人结膜上皮细胞作为诱导条件，采用分层共培养的方法诱导小鼠ESC，倒置相差显微镜观察共培养后形态的改变；免疫组织化学法检测诱导后的细胞角蛋白3／12(CK3/K12)、角蛋白13(CKl3)、MUC4和MUC5AC的表达。结果：采用人结膜上皮细胞作为诱导条件，ESC在诱导72h后可见部分细胞显示典型上皮祥细胞形态，免疫组化检测CKl3、MUC4表达阳性，CK3/K12，MUC5AC表达阴性。结论：采用分层共培养的方法，用人结膜上皮细胞作诱导剂可以诱导小鼠ESC向结膜上皮样细胞分化。眼科学报2003；19：117-121

The Study on Induced Differentiation of Embryonic Stem Cells into Conjunctival Epithelial-like Cells

PURPOSE: To investigate the probabilities of murine embryonic stem cells (ESC) differentiating into conjunctival epithelial cells and lay a basis for seeking a new source of seed cells provided in conjunctival tissue engineering. METHODS: In vitro, the ESC were induced by conjunctival epithelial cells in transwell. The morphological changes of the ESC were observed under phase contrast microscope, the specific markers of the induced cells were identified immunocytochemically with Cytokeratin 3/12 (CK3/CK12), Cytokeratin 13 (CK13), MUC4 and MUC5AC. RESULTS: The ESC might be induced by human conjunctival epithelial cells into the conjunctival epithehal-like cells after 72 hours of inducing. These cells had the typical epithelial cells morphology and were positive in cytokeratin 13, MUC4 staining, negativein CK3/CK12, MUC5AC staining. CONCLUSIONS: The murine ESC may be induced into conjunctival epithelial-like cells by conjunctival epithelial cells of human in vitro.