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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的差异,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-氮杂胞甙(5-aza)诱导CD73+/CD73-亚群,免疫荧光法检测心肌特异性肌钙蛋白T(c Tn T),Real-time PCR检测c Tn T、Gata-4变化;4’6-二脒基-2-苯基吲哚(DAPI)标记两个亚群后,移植到大鼠心肌内,免疫荧光法检测移植细胞c Tn T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 Tn T表达率为45.5%、CD73-亚群为5.75%,基因水平上CD73+亚群表达c Tn T、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 Tn T。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。
Objective To investigate the differentiation of two subpopulations of CD73 + and CD73-adipose tissue-derived mesenchymal stem cells (ADMSCs) into cardiomyocytes, and to screen out the dominant subpopulations of myocardial differentiation, providing experimental references for further cell therapy of myocardial diseases. Methods The subsets of CD73 + / CD73-ADMSCs were sorted by flow cytometry and the morphology was observed by HE staining. CD73 + / CD73- subsets were induced by 5-aza in vitro, cardiac troponin T (cTn T) was detected by immunofluorescence and cTn T and Gata-4 by Real-time PCR ; 4’6-diamidino-2-phenylindole (DAPI) labeled two subpopulations, transplanted into the myocardium of rats, immunofluorescence assay cTn T cells transplanted. Results The morphology of CD73 + ADMSCs was mainly slender or spindle-shaped in small cells. The majority of CD73- cells were flattened or square. After myocardial inducible, the expression of cTn T in CD73 + ADMSCs was 45.5% and that in CD73- subgroup was 5.75%. The levels of cTn T in CD73 + subgenome were significantly higher than those in negative subpopulations P <0.01). In vivo, CD73 + subpopulations expressed more cTn than negative ones. Conclusions CD73 + ADMSCs have a higher potential for cardiomyogenic differentiation than CD73-ADMSCs. CD73 + ADMSCs may be more promising for myocardial therapy.