低频脉冲电磁场诱导大鼠骨髓间充质干细胞软骨样细胞分化  

Low Frequence Pulsed Electromagnetic Fields Induce Chondrocyte-like Cells Differentiation of Rat Bone Marrow-derived Mesenchymal Stem Cells in Vitro

作　　者：邱飞远[1] 何学令[1] 姚晓琳[1] 李凯[1] 况薇[1] 吴文超[2] 李良[1,2] 

机构地区：[1]四川大学华西基础医学与法医学院生物医学工程学研究室,成都610041 [2]四川大学华西医院心血管疾病研究室Ⅱ,成都610041

出　　处：《生物医学工程学杂志》2012年第3期501-507,共7页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(10972149;30770534);国家重点基础研究发展计划(973)资助项目(2005CB623901)

摘　　要：骨髓间充质干细胞(BMSCs)是一类具有多向分化潜能的细胞。低频脉冲电磁场(LEPEMFs)治疗可导致哺乳动物细胞水平的生物化学变化,加速组织修复。因此,我们假设LFPEMFs可以促进体外大鼠骨髓间充质干细胞(rBMSCs)向软骨样细胞分化。实验分为3组:1)LFPEMFs组,rBMSCs暴露于50Hz、1mT低频脉冲电磁场,30min/d;2)成软骨诱导组,采用软骨诱导介质培养rBMSCs;3)对照组,采用完全培养介质培养rBMSCs。通过倒置相差显微镜逐日连续观察rBMSCs的生长状况和形态特征。分别于实验第10、15和20d收集各组rBMSCs标本,采用实时荧光定量PCR方法检测Ⅱ型胶原(ColⅡ)和聚集蛋白聚糖(aggrecan)mRNA表达,采用组织化学与免疫组化方法检测aggrecan和ColⅡ的蛋白表达。结果表明:在各时间点,LFPEMFs组与成软骨诱导组的ColⅡ、ag-grecan mRNA表达和蛋白表达明显高于对照组(P<0.05)。本实验提示,LFPEMFs可以促进rBMSCs向软骨样细胞分化。Mesenchymal stem cells（MSCs） are multipotent stem cells that differentiate into a variety of cell types. Low frequency pulsed electromagnetic fields（LFPEMFs） therapy can causes biochemical changes at the cellular level to accelerate tissue repair in mammals. So, we tested the hypothesis that LFPEMFs can promote chondrogenic dif- ferentiation of rat bone marrow-derived mesenehymal stem cells （rBMSCs） in vitro. The rBMSCs were isolated by adherence method and the third-generation of the rBMSCs were randomly divided into LFPEMFs groups, chondro- cyte-induced group and control group. LFPEMFs groups with complete medium were exposed to 50Hz, lmT PEMFs for 30 rain every day, lasting for 10, 15 and 20 d, respectively. Chondrocyte-induced group were treated with chondrogenic media, while control groups were only cultured with complete medium. The mRNA expressions of type II-collagen （Col II） and aggrecan were determined by Real-time fluorescent quantitation PCR. The protein ex- pression of Col II and aggrecan were detected with toluidine blue stain or immunocytochemical stain, respectively. The result showed that the mRNA and protein expression level of Col-II and aggrecan were significantly higher in the LFPEMFs group or chondrocyte-induced group, compared to the control group. It suggest that LFPEMFs could contribute to rBMSCs to differentiate into chondrogenic differentiation in vitro.

关 键 词：低频脉冲电磁场 大鼠骨髓间充质干细胞 二型胶原 聚集蛋白聚糖 软骨样细胞分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]