自体hcn2-mscs希氏束内移植治疗完全性房室传导阻滞的实验研究-experimental study on the treatment of complete atrioventricular block with autologous hcn 2 - mscs in his bundle.docx

下载文档 降价啦

1
0
约8.41万字
约 101页
2018-09-10 发布于上海
举报
版权申诉
保障服务

自体hcn2-mscs希氏束内移植治疗完全性房室传导阻滞的实验研究-experimental study on the treatment of complete atrioventricular block with autologous hcn 2 - mscs in his bundle.docx

1、原创力文档（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。。
2、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
3、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。

自体hcn2-mscs希氏束内移植治疗完全性房室传导阻滞的实验研究-experimental study on the treatment of complete atrioventricular block with autologous hcn 2 - mscs in his bundle

第第二军医大学博士学位论文自体 HCN2-MSCs 希氏束内移植治疗完全性房室传导阻滞的实验研究 PAGE PAGE 6 PAGE PAGE 7 Objective Abstract Biological cardiac pacemaker (BCP) is a revolutionary concept in the treatment of complete atrioventricular block. The theoretical basis of BCP is either to resume the atrioventricular (AV) conduction, or genetically altered a non-pacemaker cell to obtain a higher ventricle rate. Based on the research on mesenchymal stem cells (MSCs) in our department，this study aimed to answer the following questions of BCP： After autotransplantation into the lesion area of the His-bundle, would the MSCs bring the AV conduction back? After being implanted into severed region of the His-bundle, would the autologous pacemaker cells act with more ordered activation and contraction than they did in the free wall of ventricle? Methods Porcine MSC cultures were established following bone marrow aspiration from healthy young male pigs using gradient isolation techniques and flow cytometry. The MSCs were cultured in Dulbeccos Modified Eagle Media (DMEM) with 20% fetal bovine serum in a humidified atmosphere of 5% CO2. Cultures were maintained at subconfluence. The growth curves were made to calculate the cell doubling time； MSCs harvested were tested for expression of CD105, CD73, CD90, CD45, CD34, CD14 and CD19 by flow cytometry. The MSCs were also induced to multi-differentiate. After Ad.HCN2-GFP was constructed and titrated, the cytotoxicity was detected. The MSCs were transfected by Ad.HCN2-GFP with a MOI of different levels to determine the best one. With the best MOI, the Ad.HCN2-GFP was introduced into MSCs and then HCN2 protein was detected in the cells by methods of immunofluorescence and flow cytometry. By patch clamp, the electrophysiology of HCN2-GFP-MSCs was studied. The neonatalrat’s cardiomyocytes (NRCs) were isolated by enzyme digestion and cocultured with the transgenic MSCs. The beating rates were r