[剪切应力作用下与内皮细胞共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].

作者信息

Zhang Lin, Li Yuqnan, Zhang Chuansen, Zhang Yan, Yang Xiangqun

机构信息

Department of Anatomy, Institute of Biomedical Engineering, the Second Military Medical University, Shanghai 200433, China.

出版信息

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2009 Feb;26(1):85-8.

PMID:19334561

Abstract

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-alpha-actin (SM-alpha-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-alpha-actin. After 48 hours, they expressed SM-alpha-actin and calponin obviously. After 72 hours, obvious expressions of SM-alpha-actin and calponin, but not of SMMHC, were detected. Further static co-culture had no effect on SM-alpha-actin, calponin and SMMH expression of BMSCs; after 24 hours, shear stress induced feeble expression of SM-alpha-actin and obvious expression of SMMHC in co-cultured BMSCs, but it had no effect on the expression of calponin. The results suggest that shear stress may potentiate the differentiation of BMSCs (co-cultured with ECs) into mature smooth muscle-like cells.

摘要

研究了在剪切应力作用下与内皮细胞（ECs）共培养的骨髓间充质干细胞（BMSCs）的分化情况。将BMSCs和ECs在PET膜两侧共培养，72小时后通过平行板流动腔施加20达因/平方厘米的剪切应力。在相差显微镜下观察细胞形态，并通过荧光免疫细胞化学检测BMSCs中平滑肌α-肌动蛋白（SM-α-肌动蛋白）、钙调蛋白和平滑肌肌球蛋白重链（SMMHC）的表达。共培养的BMSCs逐渐变成平滑肌样细胞；24小时后，BMSCs开始表达SM-α-肌动蛋白。48小时后，它们明显表达SM-α-肌动蛋白和钙调蛋白。72小时后，检测到明显表达的SM-α-肌动蛋白和钙调蛋白，但未检测到SMMHC的表达。进一步的静态共培养对BMSCs的SM-α-肌动蛋白、钙调蛋白和SMMH表达没有影响；24小时后，剪切应力诱导共培养的BMSCs中SM-α-肌动蛋白的微弱表达和SMMHC的明显表达，但对钙调蛋白的表达没有影响。结果表明，剪切应力可能促进（与ECs共培养的）BMSCs向成熟平滑肌样细胞的分化。