【摘要】 目的:观察Notch信号激活剂Jagged蛋白对胚胎干细胞向心肌细胞分化的影响。方法:小鼠胚胎干细胞用荧光染料DAPI(200mg/L)标记,在细胞培养的基础上,加入Notch信号激活剂Jagged蛋白(5mg/L)作用14d。①相差显微镜下观察自发性节律收缩细胞的出现情况;②透射电镜对培养细胞进行超微结构分析;③采用免疫细胞荧光技术检测培养细胞是否有心肌特异性收缩蛋白的表达,并计算胚胎干细胞分化为心肌细胞的比例(胚胎干细胞用DAPI标记,核发蓝色荧光,心肌特异性收缩蛋白用Cy3标记的二抗检测,胞质发红色荧光)。结果:胚胎干细胞具有很强的自发分化能力,培养第14天,约15%左右的胚胎干细胞分化为具有自发性收缩活动的心肌细胞。加入Jagged蛋白(Notch信号激活剂)后,胚胎干细胞向心肌细胞分化的比例显著减少。结论:胚胎干细胞中加入Jagged蛋白持续激活Notch信号,会抑制胚胎干细胞向心肌细胞分化。更多还原

【Abstract】 Objective:To observe the effects of Jagged protein on the differentiation of embryonic stem cells to myocardial cells.Methods:Mouse embryonic stem cells were labelled by fluorescent dye DAPI(200 mg/L).Then Jagged protein(5 mg/L),the activator of notch signal,was added to embryonic stem cells in vitro for 14 days.The spontaneously contractive cells were observed under phase contrast microscope,ultra microstructure of spontaneously contractive cells were observed under transmission electron microscope.The expression of myocardial specific contactive protein was identified by cell immunofluorescent assay,and the differentiation ratio of embryonic stem cells to cardiomyocytes was calculatd.Results:The spontaneous differentiation of embryonic stem cells is strong.On the 14th day,there were about 15% embryonic stem cells differentiated into spontaneously contractive cardiomyocytes.When Jagged protein was added,the differentiation ratio was significantly reduced.Conclusion:Jagged protein added to embryonic stem cells activates the notch signal,which would inhibit the differentiation of embryonic stem cells to cardiomyocytes.更多还原