心肌细胞外基质对心肌球源性心肌干细胞体外分化、增殖及凋亡影响的研究

Effect of Rat Heart Tissue-derived Extracellular Matrix on Differentiation,Proliferation,and Apoptosis of Cardiosphere-derived Cells in Vitro

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 effect of heart tissue -derived extracellular matrix（ECM） on the differentia- tion, proliferation and apoptosis of cardiosphere-derived cells（CDC） in vitro. Methods CDCs were cultured by cardiac explant methods. ECM was prepared by decelluariztion procedure. CDCs were cultured on ECM coated dishes or conven- tional fibrin （FN） coated dishes. Then we compared the differentiation rate, proliferation, and apoptosis rate of CDC between the two groups in vitro. Results ECM could significantly promote CDC differentiating into vascular endothelial cell, cardiac muscle cell or smooth muscle cell （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）. ECM also significantly promoted the proliferation of CDC and reduced the apoptosis and necrosis rate of CDC in vitro （0.052±0.002 vs. 0.025±0.001, P〈0.001）. Conclusion We obtained c-kit CDCs, effectively remove the cellular components of heart tissue-derived ECM and preserved the composition and structure of ECM. ECM can promote the differentiation of CDC to vascular endothelial cell, cardiac muscle cell or smooth muscle cell, promote the proliferation of CDCs and decrease CDC apoptosis and necrosis rate in vitro.