Metzinger L, Poindron P, Passaquin A C
Université Louis Pasteur (ULP), Département d'Immunologie, d'Immunopharmacologie et de Pathologie, Illkirch, France.
Cytotechnology. 1993;13(1):55-60. doi: 10.1007/BF00749976.
We describe a rapid and reproducible technique for establishing primary cultures of skeletal muscle cells from mouse origin. This method was aimed at avoiding extensive enzymatic proteolysis which is commonly used for preparation of primary skeletal muscle cultures. It relies on a Stomacher blender that allows a rapid and regular mechanical dissociation of muscle samples by repeated shocks. Cultures have been compared to those obtained by a modification of the method of Yaffé (1993) based on tryptic dissociation of rat muscle thighs. The time of preparation was reduced to 1 h and 15 min as compared to 4 h with the technique of Yaffé. Both cultures displayed similar morphologies and exhibited comparable myogenesis processes. Cellular yield, rate of myotube formation and myotube numbers were similar. The expression of myogenesis markers were identical as assessed by determination of acetylcholine receptor number, creatine kinase activity and level of myosin light chain.
我们描述了一种快速且可重复的技术，用于从小鼠来源建立骨骼肌细胞的原代培养物。该方法旨在避免常用于制备原代骨骼肌培养物的广泛酶解蛋白过程。它依赖于一种均质器，通过反复冲击能使肌肉样本快速且均匀地进行机械解离。已将这些培养物与通过对Yaffé（1993年）基于胰蛋白酶解离大鼠大腿肌肉的方法进行改进而获得的培养物进行了比较。与Yaffé技术所需的4小时相比，制备时间缩短至1小时15分钟。两种培养物均呈现相似的形态，并表现出可比的肌生成过程。细胞产量、肌管形成速率和肌管数量相似。通过测定乙酰胆碱受体数量、肌酸激酶活性和肌球蛋白轻链水平评估，肌生成标志物的表达相同。
