目的 探讨CD73+和CD73-脂肪间充质干细胞（ADMSCs）两个亚群向心肌分化能力差异，筛选出心肌分化优势亚群，为进一步开展细胞治疗心肌疾患提供实验参考。方法 流式细胞仪分选CD73+/CD73- ADMSCs 亚群，HE染色观测其形态。体外5-aza诱导CD73+/CD73- 亚群，细胞免疫荧光化学法检测心肌特异性心肌肌钙蛋白c-TnT，Real-time PCR检测c-TnT、Gata-4变化；DAPI标记两个亚群后，移植到大鼠心肌内，细胞免疫荧光化学法检测移植细胞c-TnT表达。结果 CD73+ADMSCs形态以小细胞为主呈细长形或纺锤状，CD73-细胞以宽大扁平或方形等为主。体外成心肌诱导后，CD73+ADMSCs心肌特异性c-TnT表达率45.5% 、CD73-亚群5.75 %，基因水平上CD73+亚群表达c-TnT、Gata-4明显高于阴性亚群（P<0.01）。体内CD73+亚群较阴性亚群高表达c-TnT。结论 CD73+ADMSCs较CD73-ADMSCs具有更高向心肌分化的潜能，CD73+ADMSCs可能更具有心肌治疗的前景。
Abstract
Objective The cardiac differentiated potency about subpopulations of CD73+/CD73- ADMSCs were evaluated for selecting subsets of cardiac differentiation advantage. Methods Using flow cytometry to separate CD73 +/CD73- ADMSCs subgroups, their shape were detected by HE staining. After ADMSCs subgroups induced by 5-aza, the expression of c-TnT (myocardial specificity protein myocardial troponin) was detected by immunofluorescent chemical method, and using real-time PCR to check c-TnT and Gata-4 gene level changes. DAPI labeled subpopulations were myocardial transplanted, and then the expression of c-TnT were detected by immunofluorescence of transplanted cells. Results CD73+ subset were mainly small cells, possessed Stellate cells, triangle cells, slender and spindle cells. In contrast, CD73- cells showed a homogeneous morphologic pattern with wide flat or square shapes, dubbed big cells. After myocardium induction, the express rate of c-TnT is 45.5% in CD73+ ADMSCs , but in the CD73- subgroup is only 5.75%. CD73+ subsets was higher than negative subgroup in expression of c-TnT and Gata-4 gene (P < 0.01). CD73+ subsets was higher than negative subgroup in expression of c-TnT in vivo also. Conclusions The percentage of cardiomyocyte transdifferentiation in CD73+ ADMSCs was obviously higher than that in CD73- subset. The results could be demonstrated that CD73+ cell possessed preferable therapy effect for heart disease.
关键词
CD73 /
cTnT /
心肌分化 /
脂肪间充质干细胞
Key words
CD73 /
cTnT /
Cardiomyocytes differentiation /
Adipose-derived mesenchymal stem cells
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参考文献
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