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目的探讨采用小分子化合物XAV939对体外培养小鼠胚胎干细胞(mESC)诱导分化为心肌细胞的可行性。方法复苏、培养未分化的胚胎干细胞(embryonic stem cell,ESC)在小鼠胚胎成纤维细胞制作的滋养层上长至汇合,胰蛋白酶消化后经悬滴法形成拟胚体(EBs),拟胚体贴壁后用XAV939诱导分化为心肌细胞。通过光镜观察并记录跳动的心肌细胞,采用免疫荧光技术鉴定心肌细胞。结果 XAV939能有效地诱导小鼠胚胎干细胞向心肌细胞的分化,诱导平均效率为71.85%±1.05%;分化的心肌细胞自发而有节律地收缩,心肌细胞特异性标志物心肌肌钙蛋白T(cTnT)表达呈阳性。结论小分子化合物XAV939能有效地诱导小鼠胚胎干细胞在体外分化为心肌细胞。
Objective To investigate the feasibility of using small molecule compound XAV939 to induce the differentiation of mouse embryonic stem cells (mESCs) into cardiomyocytes in vitro. Methods Resuscitation and culture of undifferentiated embryonic stem cells (ESCs) were performed on confluent trophoblast cells of mouse embryonic fibroblasts. After embryos were trypsinized, embryoid bodies (EBs) were formed by hanging drop method. After XMA939 is put into the wall, the cardiomyocytes are induced to differentiate. Cardiomyocytes were observed and recorded by light microscopy. Cardiomyocytes were identified by immunofluorescence technique. Results XAV939 effectively induced the differentiation of mouse embryonic stem cells into cardiomyocytes with an average induction rate of 71.85% ± 1.05%. The differentiated cardiomyocytes contracted spontaneously and rhythmically. Cardiomyocyte-specific marker cardiac troponin T (cTnT ) Positive expression. Conclusion The small molecule compound XAV939 can effectively induce mouse embryonic stem cells to differentiate into cardiomyocytes in vitro.