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目的研究大鼠心肌组织源性细胞外基质(ECM)对心肌球源性干细胞(CDC)体外分化、增殖及凋亡的影响。方法采用心肌组织块培养法培养大鼠CDC,采用脱细胞法制备ECM,ECM包被于培养皿上,与常规CDC培养方法比较CDC体外分化、增殖及凋亡的差异。结果 ECM能够促进CDC向血管内皮细胞,心肌肌动蛋白及平滑肌细胞分化(0.060±0.002 vs.0.043±0.002,P<0.001;0.082±0.003 vs.0.051±0.002,P<0.001;0.055±0.002 vs.0.034±0.001,P<0.001);促进CDC体外增殖,降低凋亡坏死率(0.052±0.002 vs.0.025±0.001,P<0.001)。结论本实验细胞培养法可获得表达c-kit的CDC细胞,脱细胞法提取的心肌ECM可有效脱掉心肌组织中的细胞成分,较好地保留了心肌ECM的成分和结构。体外研究证实ECM可促进CDC向血管内皮、血管平滑肌、心肌细胞分化,促进增殖、降低凋亡。
Objective To investigate the effect of cardiac myogenic extracellular matrix (ECM) on the differentiation, proliferation and apoptosis of cardiac myeloid stem cells (CDC) in vitro. Methods The rat CDC was cultured with myocardial tissue block method. ECM was prepared by acellular method. ECM was coated on the culture dish. Differences of CDC in vitro differentiation, proliferation and apoptosis were compared with conventional CDC culture method. Results ECM promoted differentiation of CDC to vascular endothelial cells, cardiac actin and smooth muscle cells (0.060 ± 0.002 vs.0.043 ± 0.002, P <0.001; 0.082 ± 0.003 vs.0.051 ± 0.002, P <0.001; 0.055 ± 0.002 vs. 0.034 ± 0.001, P <0.001); promote the proliferation of CDC in vitro and reduce the necrosis rate (0.052 ± 0.002 vs.0.025 ± 0.001, P <0.001). CONCLUSION: CDC cells expressing c-kit can be obtained by the cell culture method in this experiment. The ECM extracted by acellular method can effectively remove the cellular components of cardiac tissue and retain the composition and structure of myocardial ECM. In vitro studies confirmed that ECM can promote CDC to vascular endothelial cells, vascular smooth muscle, myocardial cell differentiation, promote proliferation and reduce apoptosis.