剪切应力下与内皮细胞联合培养的骨髓间充质干细胞的分化  

Differentiation of Bone Marrow Mesenchymal Stem Cells Co-cultured with Endothelial Cells under Shear Stress

作　　者：张林[1] 李玉泉[1] 张传森[1] 张炎[1] 杨向群[1] 

机构地区：[1]第二军医大学解剖学教研室,生物医学工程研究所,上海200433

出　　处：《生物医学工程学杂志》2009年第1期85-88,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(302000053);军队“十一五”规划项目资助(06MA171)

摘　　要：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。Differentiation of bone marrow mesenchymal stem cells （BMSCs） co-cultured with endothelial cells （ECs） under shear stress was studied. BMSCs and ECs were co-cultured on the two sides of PET membrane, and 20 dyn/cm2 shear stress produced by parallel plate flow chamber was performed after 72 hours. Cell morphology was observed under phase-difference microscope, and the expressions of smooth muscle-α-actin （SM-a-actin）, calponin and smooth muscle myosin heavy chain （SMMHC） of BMSCs were detected by fluorescence immunocytochemistry. The co-cultured BMSCs became smooth muscle-like cells gradually; after 24 hours, the BMSCs started to express SM-a-actin. After 48 hours, they expressed SM-α-actin and calponin obviously. After 72 hours, obvious expressions of SM-α-actin and calponin, but not of SMMHC, were detected. Further static co-culture had no effect on SM-α-actin, calponin and SMMH expression of BMSCs; after 24 hours, shear stress induced feeble expression of SM-α-actin and obvious expression of SMMHC in co-cultured BMSCs, but it had no effect on the expression of calponin. The re- sults suggest that shear stress may potentiate the differentiation of BMSCs （co-cultured with ECs） into mature smooth muscle-like cells.

关 键 词：骨髓间充质干细胞 内皮细胞 平滑肌细胞 联合培养 剪切应力 大鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R318.01[医药卫生—基础医学]