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为探索成肌细胞增殖分化时间隙连接蛋白(connexin)43的表达和变化,建立乳鼠成肌细胞体外培养,采用0.1%胶原酶消化法分离细胞及非连续密度梯度离心法纯化细胞,以电镜及免疫组织化学法鉴定成肌细胞;采用Western blot法检测分化过程中connexin 43的变化。结果表明,成肌细胞体外培养可以表达connex- in 43,但随着细胞的融合分化,其表达量下降。结论:体外培养的成肌细胞间隙连接蛋白表达减少可能影响该细胞作为种子细胞进行心脏移植的效果。
In order to explore the expression and changes of connexin43 in myoblast proliferation and differentiation, myoblasts were established in vitro. Cells were isolated by 0.1% collagenase digestion and cells were purified by discontinuous density gradient centrifugation. The myogenic cells were identified by immunohistochemistry. The changes of connexin 43 during differentiation were detected by Western blot. The results showed that myoblasts could express connex-in 43 in vitro, but the expression of connex-in 43 decreased with the differentiation of cells. CONCLUSION: The decrease of the expression of connexin in myoblasts in vitro may affect the effect of this cell as a seed cell for heart transplantation.