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目的:探讨人骨髓间充质干细胞hMSCs(hum an m esenchym al stem cells,hMSCs)体外内皮分化基础及其诱导条件。方法:采用密度分离与贴壁筛选结合法分离hMSCs,体外联合应用生长因子VEGF165和不同细胞外基质纤维连接蛋白(FN)与Ⅰ型胶原(Col),对其进行内皮诱导分化。通过免疫细胞化学、细胞化学、流式细胞分析法及透射电镜对其分化后细胞进行鉴定。结果:hMSCs表达早期内皮分化标志之一KDR;PAS反应阳性及超微结构显示,hMSCs胞浆内含有大量糖原形成糖原池,分化后细胞胞浆外质内糖原明显减少或消失,暗示细胞发生分化;细胞诱导后CE34、β1整合素和KDR表达均增强。结论:诱导后细胞为过渡细胞群类型(transit popu lation,TP),可向内皮前体细胞(endothelial progen itor cells,EPC)EPC方向分化。推测由此类型细胞再分化为内皮细胞(endothelial cells,ECs),即hMSCs→TP→EPC→ECs。
Objective: To investigate the basis of endothelial differentiation and its induction conditions in human mesenchymal stem cells hMSCs in vitro. Methods: The hMSCs were isolated by density separation and adherent screening. VEGF165 and different fibroblasts (FN) and collagen type Ⅰ (Collagen) were used in vitro to induce endothelial differentiation. The differentiated cells were identified by immunocytochemistry, cytochemistry, flow cytometry and transmission electron microscopy. Results: KDR, one of the early signs of endothelial differentiation, was expressed in hMSCs. PAS positive reaction and ultrastructure showed that a large amount of glycogen was formed in the cytoplasm of hMSCs to form glycogen pool. The differentiation of glycogen in cytoplasm significantly decreased or disappeared Cell differentiation; CE34, β1 integrin and KDR expression were enhanced after cell induction. CONCLUSION: The induced cells are transition popu lation (TP) and can differentiate into EPCs of endothelial progenitor cells (EPCs). Speculated that this type of cell redifferentiation into endothelial cells (endothelial cells, ECs), namely hMSCs → TP → EPC → ECs.