心脏干细胞分离培养及其增殖与迁移的机制.doc

心脏干细胞分离培养及其增殖与迁移的机制 侯 波1，朱先云2，王学坤3(青岛市胶州中心医院，1心内科，2内分泌科，3青 引用本文：侯波，朱先云，王学坤. 心脏干细胞分离培养及其增殖与迁移的机制[J].中国组织工程研究，2016，20(41):6190-6196. DOI: 10.3969/j.issn.2095-4344.2016.41.017 ORCID: 0000-0002-7743-0450(侯波) 文章快速阅读： 文题释义： 干细胞nicheEdU标记增殖细胞，采用免疫荧光染色检测c-kit阳性细胞以及基质金属蛋白酶2，9及转化生长因子ββ1少量表达，但周围心肌组织上述3因子表达明显增多。④结果表明，心肌组织体外培养可获取心脏干细胞，伴随细胞增殖和迁移，其机制与基质金属蛋白酶2，9及转化生长因子β1的表达有关。 关键词： 干细胞；培养；心脏干细胞；心脏；细胞稳态；微环境；作用机制 主题词： 组织工程；干细胞；细胞增殖 Cardiac stem cells: isolation, culture, proliferation and migration Hou Bo1, Zhu Xian-yun2, Wang Xue-kun3 (1Department of Cardiology, 2Department of Endocrinology, Jiaozhou Central Hospital of Qingdao, Qingdao 266300, Shandong Province, China; 3Department of Cardiology, Qingdao Central Hospital, Qingdao 266042, Shandong Province, China) Abstract BACKGROUND: In the process of cardiac stem cell culture in vitro, the growth microenvironment may have some effects on the cell proliferation. OBJECTIVE: To investigate the possible mechanism of proliferation and migration of rat cardiac stem cells cultured in vitro. METHODS: Cardiac tissues from 10 Sprague-Dawley rats were obtained for primary culture and subculture. Passage 3 cells were collected for immunofluorescence staining, and stem cell growth factor receptor (c-kit) and CD45, CD90 were detected. Cultured tissues were collected and randomly divided into two groups: in group 1, paraformaldehyde fixation, paraffin embedding, hematoxylin-eosin staining, Masson staining, and detecting apoptotic cells using TUNEL method were conducted; in group 2, EdU labeling of proliferated cells, immunofluorescent detection of c-kit positive expression, matrix metalloproteinases 2, 9 and transforming growth factor β1 using immunofluorescent staining were done. RESULTS AND CONCLUSION: After 7-10 days of myocardial tissue culture, bright and round cells were visible, and after adhesion, fusiform cells exhibited strong growth and proliferation ability. Immunofluorescence staining showed a l