催产素诱导下ES源性心肌样细胞与原代小鼠心肌细胞特性比较研究

Comparative study on characteristics between oxytocin-induced ESC-derived cardiomyocytes and primary mouse cardiomyocytes

目的研究催产素诱导下的胚胎干细胞(embryonic stem cells,ES)源性心肌样细胞与原代小鼠心肌细胞在生理学特性及标志物表达上的差异。方法 ES细胞(S6)经复苏、传代,在体外经催产素诱导下向心肌细胞分化,诱导后第10天,分化细胞经心肌标志物α-横纹肌肌动蛋白(α-sarcomeric actin)、肌钙蛋白T(cTnT)细胞免疫荧光学染色后,荧光显微镜及光镜下观察其细胞形态和染色结果;通过Western bolt方法分析α-sarcomeric actin、cTnT、心钠素(atrial natriuretic factor,ANF)的表达情况;在电镜下观察分化细胞及小鼠原代心肌细胞的特异性结构,比较分化细胞与原代小鼠各项指标的差异。结果催产素诱导的胚胎干细胞源性心肌样细胞cTnT、α-横纹肌肌动蛋白免疫荧光染色阳性,ANF、cTnT、α-横纹肌肌动蛋白表达两组比较均无明显差异(P>0.05);电镜下观察ES源性心肌样细胞可见排列整齐的肌原纤维、肌小节结构和Z带、以及缝隙连接、桥粒连接。结论催产素诱导下ES源性心肌样细胞与原代小鼠心肌细胞在标志物表达及生理学特性上无明显差异。

Objective To compare the difference of physiological characteristics and expression of markers between oxytocin （OT）-induced ESC-derived cardiomyocytes and primary mouse cardiomyocytes （PMCs）.Methods The mESC （S6） were induced by OT to differentiate into cardiomyocytes,and then identified by staining with α-sarcomeric actin and ANF at 10 d.The specific structures of differentiated-cells and PMCs were compared under electron microscope.Western bolt was used to detect the expression of α-sarcomeric actin,cTnT,ANF in differentiated-cells and PMCs.Results The immunohisochemical results showed that α-sarcomeric actin and cardiac troponin T（cTnT） were positively expressed in oxytocin-induced ESC-derived cardiomyocytes.There was no statistically significant difference in the expression of α-sarcomeric actin,cTnT,ANF between differentiated-cells and PMCs （P ＞ 0.05）.Conclusion There is no significant difference between OT-induced ESC-derived cardiomyocytes and PMCs in physiological characteristics and expression of markers.