【作者】 朱文辉； 王予彬； 华咏； 符雪莲；

【Author】 ZHU Wen-hui~1 WANG Yu-bin~1 HUA Yong~2 FU Xue-lian~3 1.Deparment of Sports Injury and Joint Surgery,East Hospital,Tongji University,Shanghai 200120,P.R.China 2.Deparment of Central Laboratory,East Hospital,Tongji University,Shanghai 200120,P.P.China 3.Deparment of Pathology,East Hospital,Tongji University,Shanghai 200120,P.R.China)

【机构】 同济大学附属东方医院运动创伤关节外科； 同济大学附属东方医院中心实验室； 同济大学附属东方医院病理科；

【摘要】 目的探索一种体外分离纯化犬骨骼肌成肌细胞的简易方法。方法取成年比格犬股中间肌一块,采用机械分解法、一步酶消化法消化分离获取成肌细胞,利用差速贴壁法进行纯化。观察分离纯化后的成肌细胞的生长状况,绘制细胞生长曲线；并对分离纯化后的成肌细胞进行形态学研究及结蛋白免疫细胞化学的鉴定。结果采用该法分离的成年犬成肌细胞成功率高,培养过程生长旺盛,呈现出干细胞的特征,存活率高。免疫细胞化学鉴定显示 90％以上的细胞呈肌细胞特异抗体(结蛋白染色)阳性反应,说明该法获得的成肌细胞纯度较高。结论该法提供了一种体外成功分离纯化成年犬骨骼肌成肌细胞的简易方法,成功率高, 简便易行,值得推广；同时该研究也为利用犬进行成肌细胞治疗的研究奠定了实验基础。更多还原

【Abstract】 【Objective】To explore a simple procedure of the culture and purification of the canine skeletal myoblast in vitro.【Methods】The vastus intermedius of adult canine was obtained, from which myoblasts were isolated and harvested by mechanical decomposition and one-step en- zyme digestion.Then the cells were purified by combination of differential velocity adherent method. The growth state of the myoblast was observed and the myoblast growth curves were drawn;the mor- phologic properities of myoblasts were observed and myoblasts were identified by immunocytochem- istry ofdesmin.【Results】Our method had a high possibility of success in obtaining the adult ca- nine myoblasts with high survival rate in vitro and the myoblasts grew very well demonstrating char- acteristics of stem cells.More than 90% of the cells were desmin-positive by immunocytochemistry, which indicated the high purification of myoblasts.【Conclusion】Our method for isolating and puri- fying myoblasts proved to be convenient and practical with high possibility of success.At the same time,our research laid a foundation for the research of myoblast therapy by animal of canine.更多还原