人胚胎间充质干细胞向平滑肌细胞定向分化标志蛋白的选择

Selection of specific marker protein for smooth muscle lineage differentiation from human embryonic mesenchymal stem cells

目的:选择平滑肌细胞(smooth musele cell,SMC)特异性标志蛋白,作为判断人胚胎间充质干细胞(mesenchymalstem cell,MSC)是否向SMC方向分化的指标。方法:采用免疫细胞化学、免疫荧光、Western印迹和RT-PCR等方法,检测SMC分化早、中、晚期标志蛋白smooth muscle(SM)-α-actin、calponin、SM-myosin heavy chain(SM-MHC)在人胚MSC中的表达情况。结果:各实验均证明SM-α-actin和calponin蛋白在人胚MSC中表达。各种实验方法在人胚MSC中均检测不到SM-MHC蛋白及其mRNA的表达。结论:SM-MHC是一个有价值的SMC特异性标志物,可以做为判断人胚MSC向SMC方向分化与否的指标。

Objective:To select specific marker protein for smooth muscle cells (SMC) as an indicator of SMC differentiation from human embryonic mesenchymal stem cells (MSC). Methods: The expressions of smooth muscle (SM )-α-actin, calponin, and SM-myosin heavy chain (MHO , the early, middle, and late stage markers of SMC differentiation respectively, were detected in MSC by immunocytochemistry, immunofluorescence, Western blotting and RT-PCR. Results: Immunocy-tochemistry, immunofluorescence, Western blotting and RT-PCR showed that both SM-α-actin and calponin proteins expressed in human embryonic MSC. Neither protein nor mRNA of SM-MHC was found in human embryonic MSC by all 4 experiment methods. Conclusion: SM-MHC is a valuable specific marker of SMC, which can be used as an indicator of smooth muscle lineage differentiation from human embryonic MSC.