脂肪间充质干细胞CD73亚群心肌分化能力评价  

Evaluation of cardiomyocyte differentiation of CD73 subpopulations in adipose derived mesenchymal stem cells

作　　者：张珉[1,2] 高建辉[2] 李琼[2] 余霞[2] 王晶晶[2] 郭志坤[2] 

机构地区：[1]河南省肿瘤医院肝胆胰外科,郑州450008 [2]新乡医学院河南省医用组织再生重点实验室,河南新乡453003

出　　处：《解剖学报》2016年第1期49-54,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(81400237);国家大学生创新计划(201310472081)

摘　　要：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 evaluate the cardiac differentiation potency of subpopulations of CD73^＋/CD73- adlpose-derived mesenchymal stem cells （ADMSCs） for selecting advantageous subsets of cardiac differentiation. Methods Flow eytometry was used to separate CD73^＋/CD73^- ADMSCs subgroups, and their shapes were detected by HE staining. After subgroups of ADMSCs induced by 5-azacytidine （5-aza）, the expression of cardiac troponin T（cTnT） （myocardial specificity protein myocardial troponin） was detected by immunofluorescent chemical method, cTnT and Gata-4 gene level changes were checked using Real-time PCR. 4＇ 6-diamidino-2-phenylindole （DAPI） labeled subpopulations were myocardial transplanted, and then the expression of cTnT was detected by immunofluorescence of transplanted cells. Results CD73 ^＋ subsets included mainly small cells, and possessed stellate cells, triangle cells, slender and spindle cells. In contrast, CD73^- ceils showed a homogeneous morphologic pattern with wide flat or square shapes, dubbed big ceils. After myocardium induction, the express rate of cTnT was 45.5% in CD73 ^＋ ADMSCs, but only 5.75% in the CD73^- subgroup. CD73 ^＋ subsets were higher than the negative subgroup in expression of c-TnT and Gata-4 gene （P 〈 0.01 ）. CD73^＋ subsets were higher than the negative subgroup in expression of cTnT in vivo. Conclusion The percentage of eardiomyocyte transdifferentiation in CD73^ ＋ ADMSCs was obviously higher than that in CD73^- subset. The results demonstrate that CD73 ＋ cell possessed preferable therapy effect for heart diseases.

关 键 词：CD73 心肌特异性肌钙蛋白T 心肌分化 脂肪间充质干细胞 流式细胞术 小鼠 

分 类 号：Q28[生物学—细胞生物学]