大鼠皮肤成纤维细胞MyoD cDNA转染后的形态变化  

Morphological changes of dermal fibroblasts of rat following transfection with MyoD cDNA in vitro

作　　者：汪进益[1] 范慧敏[1] 刘中民[1] 丁一[1] 马亮[2] 李杨[2] 

机构地区：[1]同济大学附属东方医院胸心外科,上海200120 [2]同济大学附属东方医院细胞治疗中心,上海200120

出　　处：《同济大学学报（医学版）》2007年第1期6-9,共4页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金资助项目(30471719)

摘　　要：目的探讨运用肌肉转录调节因子(myoblast-determining,MyoD)基因转染使大鼠皮肤成纤维细胞(dermalfibroblasts,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。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.

关 键 词：皮肤成纤维细胞 肌肉转录调节因子 转基因 靶细胞 大鼠 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R394.3[医药卫生—基础医学]