体外构建血管化组织工程心肌

摘要

Obj ective :To explore the feasibility of building vascularized tissue engineering myocardium in vitro .Methods :Bone marrow mesenchymal stem cells were induced to differentiate into cardiomyocytes by 5-aza and then labeled with DAPI. Bone marrow-derived endothelial progenitor cells were induced to differentiate into endothelial cells by EGM2-MV and then labeled with CM-dil.Four million labeled cardiomyocyte cells and endothelial cells were seeded at 2 ∶ 1 ratio on Matrigel matrix glue composite as a test group,and cardiomyocyte cells were seeded onto the same kind of composite as a control group.We observed the complex morphology,cell distribution and so on.Results :In the test group where cardiomyocyte cells and endothelial cells were mixed at 2∶1 ,typical “vascular phenomenon”of endothelial cells appeared after their culture 24 h,the number of capillary-like structure was significantly higher than the control group,but myocardium apoptosis decreased compared with the control group.Conclusion:Co-culture of the bone marrow-derived endothelial progenitor cells and marrow-derived cardiac cells can successfully build vascularized tissue engineering myocardium in vitro.%目的：：探讨体外构建血管化组织工程心肌的可行性。方法：大鼠骨髓间充质干细胞以浓度为10μmol/L 的5-氮胞苷诱导为心肌细胞并用 DAPI 标记、骨髓源内皮祖细胞以内皮细胞培养基 EGM2-MV 定向诱导为内皮细胞并用 CM-dil 标记。标记后的心肌细胞与内皮祖细胞按2∶1比例，以4×106个/ml 的密度种植于 Matrigel 基质胶支架上。同种密度心肌细胞种植为对照组。对复合体形态、细胞分布等进行观察。结果：H-E 染色及荧光显微镜观察可见两组复合体的细胞分布均匀，生长状态良好；单纯心肌细胞种植组可见细胞黏附聚集生长，并未出现网状结构，而2∶1混合培养组24 h 后可见内皮细胞相互连接成网状结构，出现典型的“成血管现象”，且形成的血管样结构明显多于对照组；心肌细胞凋亡数较对照组少。结论：骨髓源性心肌细胞与骨髓源性内皮细胞联合种植于 Matrigel 基质胶共培养，可于体外成功构建血管化组织工程心肌。