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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。
Bone marrow mesenchymal stem cells (BMSCs) are a group of cells with multi-directional differentiation potential. Treatment with low-frequency pulsed electromagnetic fields (LEPEMFs) can lead to biochemical changes at the mammalian cell level, accelerating tissue repair. Therefore, we hypothesized that LFPEMFs may promote the differentiation of rat bone marrow mesenchymal stem cells (rBMSCs) into chondrocytes in vitro. The experiment was divided into 3 groups: 1) rBMSCs were exposed to 50Hz, 1mT low-frequency pulsed electromagnetic fields for 30min / d in LFPEMFs group; 2) rBMSCs were cultured in cartilage induction medium; 3) control group, rBMSCs were cultured with complete culture medium . The growth and morphological features of rBMSCs were observed daily by inverted phase contrast microscope. The rBMSCs specimens of each group were collected on the 10th, 15th and 20th day respectively. The mRNA expression of type Ⅱ collagen and aggrecan was detected by real-time fluorescence quantitative PCR. The levels of aggrecan and ColⅡ protein expression. The results showed that at each time point, the expression of ColⅡand ag-grecan mRNA and protein in LFPEMFs group and chondrocyte induced group were significantly higher than those in control group (P <0.05). This experiment suggests that LFPEMFs can promote rBMSCs to differentiate into cartilage-like cells.