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研究生理剪切应力下与内皮细胞联合培养的骨髓间充质干细胞(BMSCs)向平滑肌样细胞的分化。将BMSC与内皮细胞分别种植到PET膜两侧,静态培养72 h后给予20 dyn/cm2应力作用24 h。观察BMSCs的形态学,免疫荧光化学染色方法检测平滑肌表面标志物的表达。联合培养的BMSCs逐渐成平滑肌样细胞形态,24 h开始表达平滑肌-α-肌动蛋白(SM-α-actin),48 h明显表达钙调理蛋白(calponin),72h SM-α-actin、calponin均明显表达,而平滑肌肌球蛋白重链(SMMHC)未见表达。进一步静态培养SM-α-actin、calponin、SMMHC表达情况无变化。剪切应力作用24 h后BMSCs SM-α-actin表达减少,calponin无变化,而SMMHC明显表达。结果表明剪切应力能促进与血管内皮细胞联合培养的BMSCs向平滑肌样细胞分化。
To study the differentiation of bone marrow mesenchymal stem cells (BMSCs) co-cultured with endothelial cells to smooth muscle-like cells under physiological shear stress. The BMSC and endothelial cells were implanted on both sides of the PET film, static culture 72 d after given 20 dyn / cm2 stress for 24 h. The morphology of BMSCs was observed, and the expression of smooth muscle surface markers was detected by immunofluorescence staining. The co-cultured BMSCs gradually became smooth muscle-like cell morphology. SM-α-actin was expressed at 24 h, calponin at 48 h, SM-α-actin and calponin at 72 h However, smooth muscle myosin heavy chain (SMMHC) was not expressed. Further static culture SM-α-actin, calponin, SMMHC expression did not change. After 24 h, the expression of SM-α-actin in BMSCs decreased, calponin did not change and SMMHC was significantly expressed. The results show that shear stress can promote the differentiation of BMSCs into smooth muscle-like cells co-cultured with vascular endothelial cells.