【摘要】 目的采用简易光电成像系统记录胚胎干细胞(ES细胞)体外定向分化心肌细胞的搏动频率,为药物诱导ES细胞体外定向分化心肌细胞提供量化评价指标。方法以简易光电成像系统记录心肌细胞搏动频率,并以维A酸和淫羊藿苷诱导ES细胞定向分化心肌细胞为例,收集分化过程中发育依赖性基因(α-肌球蛋白重链、心室肌球蛋白轻链及β-肾上腺素受体)和心肌特异性蛋白(α-辅肌动蛋白和肌钙蛋白T)表达情况,同步分析光电成像信号与分子生物学指标的一致性。结果光电成像系统可灵敏反映分化心肌细胞的搏动频率,在与维A酸和淫羊藿苷共培养体系中,搏动频率与心肌发育依赖性基因、蛋白表达和β-肾上腺素受体形成等一致,可体现心肌分化的不同时间段。结论简易光电成像系统能灵敏记录ES细胞定向分化心肌细胞的搏动频率,此搏动频率与细胞分化成熟程度一致,可望成为药物诱导分化心肌细胞初步评价的量化指标。更多还原

【Abstract】 AIM To provide a simple method to investigate the inducible effects of drugs on the directional differentiation of embryonic stem (ES)cells to cardiomyocytes in vitro by adopting a photoelectricity-imaging system. METHODS Recording the beating frequency of ES cell-derived cardiomyocytes induced by retinoic acid and icariin with the photoelectricity-imaging system, to reveal the relationship between the beating frequency and the expressions of cardiac developmental-dependent genes(α-myosin heavy chain, ventricular myosin light chain and β-adrenoceptor) and cardiac-specific proteins (α-actinin, troponin T). RESULTS The beating frequency obtained from the photoelectricity-imaging system, displayed a strong correlation with the expression of cardiac developmental-dependent genes and cardiac-specific proteins during differentiation of ES cells to cardiomyocytes in vitro. CONCLUSION The beating frequency of ES cell-derived cardiomyocytes sensitively recorded with photoelectricity-imaging system displayed a positive correlation with the differentiation stages. Therefore it may offer one useful means to evaluate the inducible effects of drugs on the directional differentiation of ES cells to cardiomyocytes in vitro.更多还原