细胞间接触诱导骨髓间充质干细胞向平滑肌细胞分化的实验研究刘晓芳王延洲徐惠成梁志清(第三军医大学第一附属医院, 重庆400038).docVIP

细胞间接触诱导骨髓间充质干细胞向平滑肌细胞分化的实验研究 刘晓芳　王延洲　徐惠成　梁志清 (第三军医大学第一附属医院，重庆400038) 提要： 目的　通过两种细胞的共培养探讨细胞间接触对间充质干细胞（Mesenchymal Stem Cells, MSCs分化的影响。方法 分别采用密度梯度离心法和胶原酶消化法分离培养了骨髓间充质干细胞（Bone Marrow Mesenchymal Stem Cells ，BMSCs）The Study of Cell-Cell Contact Inducing Mesenchymal Stem Cell Differentiation to Smooth Muscle Cell Xiao-Fang Liu, Yang-Zhou Wang, Hui-Cheng Xu,Zhi-Qing Liang (The first affiliated hospital of The Third Military University, Chongqing 400038, China) Abstract: Objective To establish the co-culture system between mesenchymal stem cells （MSCs） and smooth muscle cells in vitro, and study the influence of smooth muscle cells on the differentiation of bone marrow mesenchymal stem cells (BMSCs ) into smooth muscle cells in the co-culture system. Methord We used the method of density gradient centrifugation to separate and culture BMSCs and method of Collagenase digestion to separate and culture blander smooth muscle cells, then assayed cells respectively. Afer that, We designed four group ,BMSCs alone was the positive control grope , smooth muscle cells alone was the negative control grope, BMSCs and smooth muscle cells co-culture with contact or non-contact are the two experimental group . The expression of α- SMA, calponin were detected by immunofluorescence and observed with fluorescence microscope in contact co-culture grope. The method of Western-Blot was applied to determine protein expression of calponin. Results Cells with contact co-culture induction grope were observed for the expression of calponin increase gradually. Conclusion The contact between smooth muscle cells and BMSCs can promote the differentiation of BMSCs into smooth muscle cells. Key words: cell-cell contact, microenvironment, mesenchymal stem cell, smooth muscle cell 尿失禁是女性的常见病，据报道此病的患病率在10% －50%1,2 ,其中又以压力性尿失禁为主 ,已成为重要的妇女健康问题，影响生活质量，现有的研究表明其主要的病因与尿道周围的横纹肌和平滑肌的病变或凋亡相关， 近年来干细胞移植成为肌肉组织损伤修复的重要手段3 ,亦为尿失禁的治疗提供了一种新的途径4,5。骨髓间充质干细胞（Bone Marrow Mesenchymal Stem Cells,BMSCs）由于其多向分化潜能，以及低免疫原