利用c-kit+骨髓间充质干细胞与心脏脱细胞骨架共培育心肌组织

利用c-kit+骨髓间充质干细胞与心脏脱细胞骨架共培育心肌组织
张光伟，谷天祥，管晓宇，张玉海
中国医科大学附属第一医院心脏外科，辽宁省沈阳市110001
Cultivation of myocardial tissue by using c-kit+ bone marrow mesenchymal stem cells and decellularized heart matrix
Zhang Guang-wei, Gu Tian-xiang, Guan Xiao-yu, Zhang Yu-hai
Department of Cardiac Surgery, the First Hospital of China Medical University, Shenyang 110003, Liaoning Province, China
摘要
背景：研究发现c-kit阳性骨髓间充质干细胞可特异分化为心肌细胞，可能成为理想的种子细胞。

目的：验证利用c-kit+骨髓间充质干细胞及心脏脱细胞骨架培育心肌组织的可行性。

方法：取成年大鼠心脏脱细胞后备用。

取大鼠骨髓间充质干细胞，培养至第8代后行c-kit富集，采用5 μmol/L的5氮杂胞苷诱导2周后行二氢吡啶受体α2富集，然后继续培养行心肌分化鉴定或种植于心脏脱细胞骨架内行心肌组织培养。

6周后，利用心肌细胞特异性蛋白表达及动作电位鉴定心肌细胞分化，利用免疫荧光染色分析新生心肌组织结构。

结果与结论：第2次富集6周后，约60%的骨髓间充质干细胞表达心肌肌钙蛋白T、GATA结合蛋白4、连接蛋白43，电生理分析显示这些细胞可产生心肌细胞样动作电位，证实c-kit+骨髓间充质干细胞可特异分化为心肌细胞，种植于脱细胞心脏骨架后可形成排列有序的心肌组织。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
全文链接：
关键词：干细胞；骨髓干细胞；骨髓间充质干细胞；心肌细胞；脱细胞心脏；c-kit+；组织工程心脏；
Abstract：
BACKGROUND: Studies have found that c-kit+ bone marrow mesenchymal stem cells can differentiate into myocardial cells specifically, which may be the ideal seed cells.
OBJECTIVE: To investigate the feasibility of cultivation of myocardial tissue by using c-kit+ bone marrow mesenchymal stem cells and decellularized heart matrix.
METHODS: Heart tissues harvested from adult rats were decellularized for the following experiments. Primary rat bone marrow-derived mesenchymal stem cells were cultured in vitro. Until passage 8, bone marrow mesenchymal stem cells were enriched for c-kit and induced by 5 μmol/L 5-azacytidine for 2 weeks, and a second enrichment for the dihydropyridine receptor subunit α2δ1 was performed before analysis of cardiac differentiation or implantation into decellularized heart matrix for cultivation of myocardial tissue. Six weeks later, myocardial differentiation was identified by specific cardiac protein and action potential. Immunofluorescence staining was used to analysis neonatal myocardial tissue.
RESULTS AND CONCLUSION: Six weeks after the second enrichment, 60% bone marrow mesenchymal stem cells expressed cardiac troponin T, GATA binding protein 4, and connexin 43, and these cells could be induced to yield cardiac action potential, which was identified as cardiac differentiation. And when implanted into decellularized heart matrix, these cells could form myocardial tissue arranged regularly.
中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血。