Yan Peng-ke, Duan Cai-wen, Li Shi-huang, Mei Zheng-rong, Situ Bing, Xiao Guo-hong
The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou 510150, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2010 May;30(5):989-92.
To establish the model of bone mesenchymal stem cell-derived smooth muscle cells (BMSC-SMCs) and investigate the role of BMSC-SMCs in the development and progression of artherosclerosis.
BMSCs were isolated from the femoral bone of SD rats by adherent tissue culture method, and vascular smooth muscle cells (VSMCs) were obtained from the thoracic aorta. The differentiation of BMSCs into BMSC-SMCs was induced in the conditioned medium. The specific markers of BMSCs and BMSC-SMCs were identified by immunofluorescence (IF) staining. After treatment with 80 mg/L oxidative low-density lipoprotein (ox-LDL) for 72 h, the growth characteristics of BMSC-SMCs and VSMCs were observed. Flow cytometry was applied to analyze the cell cycle of BMSC-SMCs and VSMCs.
BMCS-SMCs transformed into foam cells after treatment with ox-LDL, which was more obvious in comparison with VSMCs. The growth curve of BMSC-SMCs and VSMCs presented with an S-shape pattern with the cell doubling time of 20 and 32 h, which was reduced to 15 and 28 h after treatment with 80 mg/L ox-LDL, respectively. Flow cytometry showed that exposure to 80 mg/L ox-LDL significantly increased G(0)/G(1) and decreased S and G(2)/M phase cells in both BMSC-SMCs (P<0.01, n=3) and VSMCs (P<0.05, n=3) in comparison with the control cells.
BMSC-SMC might be involved in the formation of fatty core and accelerate the development of atherosclerosis.
建立骨间充质干细胞来源的平滑肌细胞(BMSC-SMCs)模型,并研究BMSC-SMCs在动脉粥样硬化发生发展中的作用。
采用贴壁组织培养法从SD大鼠股骨中分离骨髓间充质干细胞(BMSCs),从胸主动脉获取血管平滑肌细胞(VSMCs)。在条件培养基中诱导BMSCs分化为BMSC-SMCs。通过免疫荧光(IF)染色鉴定BMSCs和BMSC-SMCs的特异性标志物。用80mg/L氧化型低密度脂蛋白(ox-LDL)处理72h后,观察BMSC-SMCs和VSMCs的生长特性。应用流式细胞术分析BMSC-SMCs和VSMCs的细胞周期。
用ox-LDL处理后,BMSC-SMCs转变为泡沫细胞,与VSMCs相比更明显。BMSC-SMCs和VSMCs的生长曲线呈S形,细胞倍增时间分别为20和32h,用80mg/L ox-LDL处理后分别降至15和28h。流式细胞术显示,与对照细胞相比,暴露于80mg/L ox-LDL显著增加了BMSC-SMCs(P<0.01,n=3)和VSMCs(P<0.05,n=3)中的G(0)/G(1)期细胞,减少了S期和G(2)/M期细胞。
BMSC-SMC可能参与脂肪核的形成并加速动脉粥样硬化的发展。