人脐带间充质干细胞向心肌样细胞分化的研究

发布时间：2018-05-20 02:29

本文选题：脐带 + 间充质干细胞　；参考：《汕头大学》2007年硕士论文

【摘要】： 目的：探讨人脐带间充质干细胞(MSCs)向心肌细胞分化的可能性，为心肌细胞的移植探索新的细胞来源。方法：从人脐带华尔通氏胶分离、培养MSCs，，用(5-azacytidine，5-aza)和二甲基亚砜(DMSO)诱导人脐带间充质干细胞的分化。观察诱导后分化细胞形态的变化，用免疫细胞化学方法检测分化的细胞是否表达心肌肌钙蛋白I(troponin I)、心肌肌钙蛋白T(troponinT)和心肌结蛋白(desmin)，RT-PCR方法测定分化的细胞是否有心肌特异转录因子(Nkx2.5)和心肌细胞结蛋白(desmin)的cDNA的表达。透射电镜观察分化的心肌样细胞内是否有肌丝的存在，扫描电镜观察分化的心肌样细胞的形态变化，从不同的层面鉴定诱导分化的心肌样细胞是否具有心肌细胞的特性和功能? 结果：从人脐带华尔通胶分离、培养的贴壁细胞，体外生长形态类似于成纤维细胞，可以维持在未分化状态稳定增殖，体外增殖超过10代，细胞冻存一个月后复苏，生长特点与冻存前基本一致。人脐带MSCs经5-aza和DMSO诱导后可向心肌样细胞分化，分化的细胞表达心肌细胞的标记troponin I、troponinT和desmin。RT-PCR检测证实，人脐带MSCs诱导前不表达Nkx2.5和desmin，经5-aza和DMSO诱导后，分化的细胞检测到有心肌特异转录因子(Nkx2.5)和心肌细胞结蛋白(desmin)的cDNA的表达。透射电镜可以观察分化的细胞浆中有肌丝的存在。结论：人脐带间充质干细胞可以分化为心肌样细胞，5-aza和DMSO可以作为心肌细胞诱导剂，人脐带间充质干细胞可以作为心肌细胞的来源。
[Abstract]:Aim: to explore the possibility of differentiation of human umbilical cord mesenchymal stem cells (MSCs) into cardiomyocytes and explore a new source of cells for cardiac myocyte transplantation. Methods: human umbilical cord mesenchymal stem cells (MSCs) were isolated from human umbilical cord and cultured with 5-azacytidine (5-aza) and dimethyl sulfoxide (DMSO) to induce the differentiation of human umbilical cord mesenchymal stem cells. The morphological changes of differentiated cells were observed after induction. Immunocytochemistry was used to detect whether differentiated cells expressed cardiac troponin I(troponin, cardiac troponin T) and cardiac desmin. RT-PCR method was used to detect whether differentiated cells had myocardial specific transcription factor Nkx2.5) and cardiomyocyte egg formation. The expression of cDNA. Transmission electron microscopy (TEM) was used to observe the presence of myofilament in differentiated cardiomyoid cells. SEM was used to observe the morphological changes of differentiated cardiomyoid cells and to identify whether the differentiated cardiomyocyte-like cells had the characteristics and functions of cardiomyocytes from different levels. Results: the adherent cells were isolated from human umbilical cord and cultured in vitro. The growth morphology of adherent cells was similar to that of fibroblasts. The cells could proliferate steadily in undifferentiated state and proliferate in vitro for more than 10 generations. The cells were resuscitated after one month of cryopreservation. The growth characteristics were basically consistent with those before freezing. Human umbilical cord MSCs could differentiate into cardiomyocyte-like cells induced by 5-aza and DMSO. The detection of troponin Itropon T and desmin.RT-PCR showed that human cord MSCs did not express Nkx2.5 and desmin before induction, but 5-aza and DMSO induced them. The expression of cDNA was detected in differentiated cells with myocardial specific transcription factor Nkx2.5) and cardiac myocyte desmin. The presence of myofilms in the differentiated cytoplasm can be observed by transmission electron microscopy (TEM). Conclusion: human umbilical cord mesenchymal stem cells can differentiate into cardiomyocyte-like cells, 5-aza and DMSO can be used as inducers of cardiomyocytes, and human umbilical cord mesenchymal stem cells can be used as sources of cardiomyocytes.
【学位授予单位】：汕头大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

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