心肌营养素1诱导骨髓间充质干细胞分化为心肌样细胞  

Differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells in vitro via cardiotrophin 1

作　　者：彭朝权[1] 高雅[1] 项鹏[2] 杨珂[1] 邹丽媛[1] 吴晓[1] 

机构地区：[1]中山大学附属第三医院心内科,广东广州510630 [2]中山大学干细胞与组织工程研究中心,广东广州510080

出　　处：《中国病理生理杂志》2011年第11期2061-2066,共6页Chinese Journal of Pathophysiology

基　　金：广东省科技厅资助项目(No.2009B030801133)

摘　　要：目的:探索心肌营养素1(CT-1)对小型猪骨髓间充质干细胞(MSCs)向心肌样细胞分化的影响。方法:获取、扩增稳定的成体西藏小型猪MSCs,鉴定成脂、成骨潜能。分4组诱导向心肌样细胞分化,包括空白对照组、5-氮杂胞苷(5-Aza)组、CT-1组、5-Aza和CT-1合用组。取诱导后4周的细胞加肌动蛋白(α-actin)和心肌肌钙蛋白T(cTnT)抗体,进行免疫荧光染色,最后计数红色荧光阳性染色率。结果:分组诱导分化结果,合用组的诱导分化心肌样细胞α-actin阳性率为29.90%±4.76%,明显大于5-Aza组(17.73%±2.35%,P<0.01)、CT-1组(6.63%±0.55%,P<0.01)和空白对照组(1.62%±0.09%,P<0.01);5-Aza组明显大于CT-1组(P<0.01)和空白对照组(P<0.01);CT-1组明显大于空白对照组(P<0.05)。cTnT红色荧光染色结果显示:合用组的诱导分化心肌样细胞cTnT阳性率为36.50%±4.09%,明显大于5-Aza组(14.37%±1.65%,P<0.01)、CT-1组(7.50%±0.61%,P<0.01)和空白对照组(1.12%±0.23%,P<0.01);5-Aza组明显大于CT-1组(P<0.01)和空白对照组(P<0.01);CT-1组明显大于空白对照组(P<0.01)。结论:适宜浓度的5-Aza(10μmol/L)和CT-1(0.1μg/L)能够在体外诱导小型藏猪骨髓MSCs转化为心肌样细胞,诱导后的细胞获得部分心肌细胞特异性蛋白的表达。CT-1与5-Aza联合可明显提高诱导率。AIM： To investigate the effects of cardiotrophin 1（CT-1） on differentiation of swine bone marrow mesenchymal stem cells（MSCs） into cardiomyocyte-like cells in vitro.METHODS： MSCs were isolated and proliferated from Tibet miniswine.Adipogenic and osteogenic potentials were identified.MSCs were divided into 4 groups for induction： untreated group,5-azacytidine（5-Aza） group,CT-1 group and 5-Aza combined with CT-1 group.After induction for 4 weeks,the expression of cardiac cell markers including α-actin and cardiac troponin-T（cTnT） was estimated by immunofluorescence staining.Finally,the rates of red fluorescence positive-staining cells were calculated.RESULTS： The expression of α-actin in the 4 groups by red fluorescence staining was as follows： the differentiation rate of cardiomyocyte-like cells in combination group was 29.90%±4.76%,significantly higher than that in 5-Aza group（17.73%±2.34%,P0.01）,CT-1 group（6.63%±0.55%,P0.01） and untreated group（1.62%±0.09%,P0.01）.The differentiation rate in 5-Aza group was significantly higher than that in CT-1 group（P0.01） and untreated group（P0.05）.The differentiation rate in CT-1 group was significantly higher than that in untreated group（P0.01）.The expression of cTnT in the 4 groups was as follows： the differentiation rate of cardiomyocyte-like cells in combination group was 36.50%±4.09%,significantly higher than that in 5-Aza group（14.37%±1.65%,P0.01）,CT-1 group（7.50%±0.61%,P0.01） and untreated group（1.12%±0.23%,P0.01）.The differentiation rate in 5-Aza group was significantly higher than that in CT-1 group（P0.01） and untreated group（P0.01）.The differentiation rate in CT-1 group was significantly higher than that in untreated group（P0.01）.CONCLUSION： Appropriate concentrations of 5-Aza（10 μmol/L） and CT-1（0.1 μg/L） induce swine bone marrow MSCs to differentiate into cardiomyocyte-like cells in vitro.CT-1 combined with 5-Aza significantly increases the differentiation rate.

关 键 词：心肌营养素1 间充质干细胞 心肌样细胞 

分 类 号：R541.4[医药卫生—心血管疾病]