Musarò Antonio, Barberi Laura
Department of Histology and Medical Embryology, Istituto Pasteur-Fondazione Cenci Bolognetti, Interuniversity Institute of Myology, Sapienza University of Rome, Rome 00161, Italy.
Methods Mol Biol. 2010;633:101-11. doi: 10.1007/978-1-59745-019-5_8.
Muscle tissue culture provides a system for studying the growth and differentiation of muscle cells in a controlled environment. In mature muscle tissue, terminally differentiated myocytes form multinucleate syncytia in which structural and regulatory genes are expressed and the contractile apparatus is assembled. Adult muscle fibres are characterized by the presence of satellite cells. These are a quiescent population of myogenic cells that reside between the basal lamina and the plasmalemma of terminally differentiated muscle fibres and are rapidly activated in response to appropriate stimuli. This chapter describes protocols used in our laboratory for isolating and culturing satellite cells isolated from mouse skeletal muscles. In particular we discuss the technical aspect of satellite cell isolation, the methods necessary to enrich the satellite cell fraction, and the culture conditions which optimize proliferation and myotube formation of mouse satellite cells.
肌肉组织培养提供了一个在可控环境中研究肌肉细胞生长和分化的系统。在成熟的肌肉组织中，终末分化的肌细胞形成多核合胞体，其中结构和调节基因得以表达，收缩装置得以组装。成年肌纤维的特征是存在卫星细胞。卫星细胞是一群静止的生肌细胞，位于终末分化肌纤维的基膜和质膜之间，并在受到适当刺激时迅速被激活。本章描述了我们实验室用于从小鼠骨骼肌中分离和培养卫星细胞的方案。特别是，我们讨论了卫星细胞分离的技术方面、富集卫星细胞组分所需的方法，以及优化小鼠卫星细胞增殖和肌管形成的培养条件。
