小鼠成肌细胞的培养纯化和鉴定方法  

Identification, Purification and Culture of Mouse Skeletal Muscle Cell

作　　者：王训[1] 张成[1] 刘焯霖[1] 陈国俊[1] 

机构地区：[1]中山医科大学附属第一医院神经内科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2000年第S1期6-9,59,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金!资助项目 (39870 80 4);广东省自然科学基金!资助项目 (970 0 6 1);卫生部临床学科重点攻关项目!(970 40 2 2 9)

摘　　要：【目的】建立体外成肌细胞分离培养、纯化和鉴定方法。【方法】 40只小鼠分 10次采用分离消化法培养成肌细胞 ,分别用A、B、C、D 4组不同培养液培养 ,以Ficoll梯度液纯化 ,进行Desmin免疫组化鉴定。【结果】成肌细胞培养后经台盼蓝检查成活率达 95 %以上 ,在接种后 4～ 5d增殖达高峰 ,A、B、C 3组培养液对成肌细胞的培养效果无差异。Ficoll分离纯化的成肌细胞纯度较高 ,有 97%的成肌细胞Desmin胞浆呈阳性反应。【结果】采用Ficoll纯化以国产血清培养 ,可获得高产量高纯度的成肌细胞。Objective To establish a practical methods of the culture, purification and identification of mouse myoblasts in vitro. Methods Muscle samples of 40 mice were minced and digested with trypsin, cultured with four kinds of media namely A, B, C, and D, finally purified with Ficoll gradient liquid. Myogenesis were dynamically observed, myoblast were tested by Desmin immunohistochemistry stains. Result The myoblast cell viability is up to 95% by Trypan blue, the cell proliferation arrived climax during 4～5 day after planted. There is no difference between A, B, and C culture liquid. The cell purity is up to 97% by Desmin immunohistochemical stains test. Conclusion Myoblasts could be purified by Ficoll and cultured with DMEM culture liquid contained serum made in our country, in high quantity.

关 键 词：成肌细胞 细胞培养 纯化 免疫细胞化学 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]