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目的：用胚胎干细胞（ＥＳＣ）体外诱导分化为心肌细胞，从分子水平上研究早期心脏发育相关基 因及其功能。方法：（１）胚胎干细胞的培养。（２）胚胎于细胞的分化培养。（３）被分化的心肌细胞鉴定：ＲＮＡ的 提取；心脏特异性引物的合成和ＲＴ－ＰＣＲ反应；探针标记、纯化和比放射活性测定；ＲＮＡ斑点杂交。结果：用最 适条件培养液对ＦＳＣ定向诱导分化，可使８０％以上的ＥＳＣ分化为心肌细胞。心肌细胞以同步的方式进行收缩。 反转录ＰＣＲ和斑点杂交的结果显示：心肌在早期发育就开始表达其特异性基因。结论：胚胎干细胞体外能诱导 分化为心肌细胞。胎牛血清、二甲基亚砜和维甲酸的浓度及它们之间的组成不同，对ＥＳ细胞定向诱导为心肌 细胞均有影响。最佳诱导条件是用２ｎｍｏｌ／Ｌ维甲酸、０．６％二甲基亚砜和２０％胎牛血清组成的条件培养液。
OBJECTIVE: To differentiate cardiomyocytes from embryonic stem cells (ESC) in vitro and to study the genes related to early cardiac development and their functions at the molecular level. Methods: (1) Embryonic stem cell culture. (2) embryo in the cell differentiation culture. (3) Differentiated cardiomyocyte identification: RNA extraction; cardiac-specific primer synthesis and RT-PCR reactions; probe labeling, purification and specific radioactivity assay; RNA dot blot hybridization. Results: With optimal conditioned medium for directional differentiation of FSC, more than 80% of ESC could be differentiated into cardiomyocytes. Cardiomyocytes contract in a synchronized manner. The results of reverse transcription PCR and dot blot showed that the myocardium began to express its specific genes in early development. Conclusion: Embryonic stem cells can differentiate into cardiomyocytes in vitro. Fetal calf serum, dimethyl sulfoxide and retinoic acid concentrations and the composition of the difference between them, the ES cell-directed induction of myocardial cells are affected. The best induction conditions were conditioned medium consisting of 2 nmol / L retinoic acid, 0.6% dimethylsulfoxide and 20% fetal bovine serum.