现代医药卫生2017，Vol.33Issue(3)：337-339,3.DOI:10.3969/j.issn.1009-5519.2017.03.007
小鼠原代成肌细胞分离、纯化及培养
Isolation,purification and culture of primary mouse myoblast
摘要
Abstract
Objective To establish the methods of isolation,purification and culture of primary mouse myoblast,then to induce its myogenic differentiation and to detect the expression of microRNA-1(miRNA-1) and miRNA-155 during this process. Methods The primary mouse myoblast was isolated and purified from the hind leg of neonatal mouse with collagenase digestion , and difference of cell adherence. Its myogenic differentiation was induced by the differentiation culture medium containing horse serum. The myogenic ability of primary myoblast was identified by myosin heavy chain antibody (MF20) immunofluorescence staining. The expression of miRNA-1 and miRNA-155 during the myogenic differentiation was detected by Taqman real time PCR. Result The primary mouse myoblast was successfully isolated and purified;the myogenic differentiation of primary myoblast was successfully induced and MF20 immunofluorescence staining was positive. The miRNA-1 expression was dramatically up-regulated, while the miRNA-155 expression was down-regulated during the myogenic differentiation of primary mouse myoblast ,the difference was statistically significant(P<0.05). Conculsion The isolation and purification method of primary mouse myoblast is successfully constructed by this study ,which provides a model in vitro for investigating the miRNA gene regulation during myo-genic differentiation process.关键词
成肌细胞/细胞,培养的/细胞分化/微RNAsKey words
Myoblasts/Cells,cultured/Cell differentiation/MicroRNAs引用本文复制引用
熊雷,何衍佶,戴威,赵圆,高彦飞,邓忠良,聂茂..小鼠原代成肌细胞分离、纯化及培养[J].现代医药卫生,2017,33(3):337-339,3.基金项目
国家自然科学基金资助项目（81501867）。 （）
