骨髓间充质干细胞分化为心肌样细胞相关基因表达与形态结构发生的关联

发布时间：2018-05-26 08:30

本文选题：间质干细胞 + 流式细胞术　；参考：《中国组织工程研究》2017年13期

【摘要】：背景:前期已完成体外定向诱导骨髓间充质干细胞分化为心肌样细胞的实验。目的:观察骨髓间充质干细胞分化为心肌样细胞过程中,细胞形态变化、结构发生规律与GATA-4、Nkx-2.5和a-MHC表达的关联性。方法:采用心肌组织裂解液诱导SD乳鼠骨髓间充质干细胞分化为心肌细胞,以免疫组织化学染色检测心肌肌钙蛋白T和间隙连接蛋白43表达情况,鉴定心肌细胞;在定向诱导分化过程中,RT-PCR检测GATA-4、Nkx2.5与α-MHC的表达情况。结果与结论:(1)细胞形态:在定向诱导过程中,细胞形态由长梭形经历短杆状、形成短小突起、网状结构、呈竹节状及肌管样结构;(2)心肌细胞鉴定:竹节样结构开始出现心肌肌钙蛋白T和间隙连接蛋白43阳性细胞,肌管样结构心肌肌钙蛋白T和间隙连接蛋白43阳性细胞增多;(3)RT-PCR检测:在定向分化过程中,细胞GATA-4、Nkx2.5与α-MHC基因表达持续增加;细胞交错连接成网状时,GATA-4表达明显增加,之后持续高表达;相邻细胞融合形成肌管样结构时,α-MHC表达达到峰值;Nkx2.5表达随诱导时间的延长逐渐增加;(4)结果表明:骨髓间充质干细胞分化为心肌样细胞过程中,心肌细胞特异基因的表达与心肌样细胞的形态特征、特殊结构发生密切相关,具有典型的时序性和空间性。
[Abstract]:Background: the in vitro induction of bone marrow mesenchymal stem cells into cardiomyocyte-like cells has been completed. Aim: to investigate the relationship between the morphological changes of bone marrow mesenchymal stem cells (BMSCs) and the expression of GATA-4nkx-2.5 and a-MHC during differentiation into cardiomyocyte-like cells. Methods: bone marrow mesenchymal stem cells from neonatal SD rats were induced to differentiate into cardiomyocytes by myocardial tissue lysate. The expression of cardiac troponin T and gap junction protein 43 were detected by immunohistochemical staining to identify cardiac myocytes. The expression of GATA-4, Nkx2.5 and 伪 -MHC was detected by RT-PCR during directed differentiation. Results and conclusion: in the process of directional induction, the cell morphology was from long fusiform to short rod shape, forming short protuberance and reticular structure. Cardiac myocyte identification: cardiac troponin T and gap junction 43 positive cells began to appear in bamboo gangliform structure. The expression of GATA-4Nkx2.5 and 伪 -MHC gene in cardiac troponin T and interstitial connexin 43 positive cells was continuously increased during the directional differentiation, and the expression of GATA-4 was significantly increased when the cells were interlaced into reticulation. The expression of 伪 -MHC increased gradually with the prolongation of induction time. The results showed that during the differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells, the expression of 伪 -MHC reached a peak when the adjacent cells fused to form myocyte-like structure, and the expression of Nkx2.5 increased gradually with the prolongation of induction time. The expression of cardiomyocyte specific genes is closely related to the morphologic characteristics and special structure of cardiomyoid cells, and has typical temporal and spatial characteristics.
【作者单位】：广西医科大学组织学与胚胎学教研室;
【基金】：广西急诊与医学救援人才小高地·广西高校急诊医学重点实验室开放课题(GXJZ201505) 广西医科大学青年科学基金资助项目(GXMUYSF201528) 广西自然科学基金(2012GXNSFAA053126,桂科自0832157,桂科攻0592007-1G,桂科青0229009) 广西高校人体发育与疾病研究重点实验(0102402214009C)~~
【分类号】：R329.2

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