无血清EGM-2 BulletKit对骨髓间充质干细胞向血管内皮细胞分化的诱导作用
摘要
目的探讨无血清内皮细胞生长培养基套装(EGM-2 Bullet Kit)对骨髓间充质干细胞(MSC)向血管内皮细胞分化的诱导作用。方法通过贴壁培养获取MSC,用流式细胞仪检测MSC表面标志物表达。将获得的MSC随机分为对照组与诱导组。对照组用无血清EGM-2基础培养基进行培养,诱导组选用无血清EGM-2基础培养基+Bullet Kit进行诱导。于诱导第2、6、10、14天用流式细胞仪、细胞免疫荧光法检测内皮细胞表面分子CD31表达。采用Di I标记的乙酰化低密度脂蛋白吞噬实验检测MSC诱导分化后的吞噬功能,用基底膜基质胶成管实验检测细胞体外成管功能。结果贴壁培养获得的MSC呈长梭形,流式细胞仪检测结果显示MSC标志物表达阳性的比例均大于95%,而内皮细胞标志物无表达。诱导组诱导第2天CD31开始表达,随诱导时间增加,CD31表达量逐渐升高,诱导第14天CD31表达量最高,CD31表达阳性的内皮细胞比例为55.8%;对照组无CD31表达。诱导第14天,诱导组能够摄取脂质,而对照组无摄取现象。诱导组细胞能够形成典型的管状结构;对照组细胞未形成典型的线状、管状结构。结论无血清EGM-2 Bullet Kit可诱导MSC分化为血管内皮细胞,且具有内皮细胞的特性、功能。
引文
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[5]Shimamura M,Nakagami H,Koriyama H,et al.Gene therapy and cell-based therapies for therapeutic angiogenesis in peripheral artery disease[J].Biomed Res Int,2013,2013:186215.
[6]张凯,王毅,邢国胜.间充质干细胞的生物学特性及多向分化潜能[J].中国组织工程研究与临床康复,2008,12(3):539-542.
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[9]Xu ZC,Zhang Q,Li H.Differentiation of human hair follicle stem cells into endothelial cells induced by vascular endothelial and basic fibroblast growth factors[J].Mol Med Rep,2014,9(1):204-210.
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