中文摘要：

目的 探讨运用肌肉转录调节因子（myoblast-de termining,MyoD）基因转染使大鼠皮肤成纤维细胞（dermal fibroblasts,DFs）转化为肌细胞的可能性.方法 采用差速贴壁法培养Sprague-Dawley（SD）大鼠的DFs,利用携带MyoD cDNA的真核表达载体pLenti6/V5- DEST-MyoD转染培养的大鼠DFs,经Blasticidin 筛选2周后挑选转基因单克隆细胞,传代培养,进行细胞形态学观察及细胞免疫化学鉴定.结果 大鼠DFs在导入MyoD cDNA后可见：（1） 部分细胞形态发生改变, 类似骨骼肌细胞,胞体伸展,胞浆颗粒增多,（2） 经抗Desmin抗体的细胞免疫化学检测证实转染后的细胞有下游蛋白Desmin的阳性表达. 结论 DFs能转化为具有潜在功能的类骨骼肌细胞,提示DFs可成为心血管疾病基因治疗的靶细胞.

英文摘要：

Objective To evaluate the possibility of converting cultured rat dermal fibroblasts（DFs） to skeletal muscle cells by transfection with myoblast-determining gene（MyoD cDNA）,one of the basic helix-loop-helix myogenic factors.Methods DFs of Sprague-Dawley（SD） rat were isolated and cultured with an improved technique of differential anchoring velocity,MyoD cDNA was transduced into DFs by using the eukaryotic expression vector pLenti6/V5-DEST-MyoD,and the cells were incubated in medium containing Blasticidin for screening for 2 weeks. Resistant colonies were picked and subcultured until use. The morphologic characters of those cells were examined by inverted microscope and immunocytochemical staining. Results The DFs transletted with MyoD cDNA showed that： （ 1 ） Some of them were morphologically similar to skeletal muscle cells, cell body elongated and cytoplasmic particles increased; （2） The expression of downstream myogenic Desmin protein （Muscle cell marker） was observed in the transfected cells with immunocytochemical staining. Conclusion The DFs can be converted to＂ skeletal muscle-like＂ cell with potential function. These results suggest that DFs could be one of the ideal target cells for genetic therapy of molecular cardiomyoplasty.