ISOLATION AND CHARACTERIZATION OF HUMAN-MUSCLE CELLS
ISOLATION AND CHARACTERIZATION OF HUMAN-MUSCLE CELLS
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DOI：
10.1073/pnas.78.9.5623
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
WEBSTER, C
中科院分区：
文献类型：
--
作者：
BLAU, HM;WEBSTER, C
描述了一种在标准化条件下研究出生后人体肌肉的体外系统。该技术利用克隆来分离肌肉细胞的纯群体。通过操纵培养条件，可以使单个克隆或合并的克隆的增殖或分化最大化，以产生肌细胞的大量培养物。肌肉表型稳定;细胞可以储存在液氮中长期使用，而不会丧失增殖或分化潜力。肌细胞的分化能力由克隆生长动力学分析确定;分化能力由形态学证据（细胞融合、条纹、收缩和乙酰胆碱受体的出现）和肌肉蛋白质合成的生化分析（肌酸激酶，α-磷酸肌酸激酶，α-磷酸肌酸激酶）确定。肌动蛋白原肌球蛋白和肌球蛋白L链）。这种方法消除了迄今为止使初级肌肉研究复杂化的细胞组成的可变性。可以以受控的方式研究不同细胞类型对正常和遗传性营养不良的人类肌肉发育的相互作用和贡献。
An in vitro system for the study of postnatal human muscle under standardized conditions is described. The technique utilizes cloning to isolate pure populations of muscle cells. By manipulating culture conditions either proliferation or differentiation of individual clones or of clones pooled can be maximized to yield mass cultures of muscle cells. The muscle phenotype is stable; cells can be stored in liquid N for long-term use without loss of proliferative or differentiative potential. Proliferative capacity of muscle cells was determined from an analysis of clonal growth kinetics; differentiative capacity was determined from morphological evidence (cell fusion, striations, contractions and the appearance of acetylcholine receptors) and biochemical analysis of muscle protein synthesis (creatine kinase, .alpha.-actin tropomyosin and myosin L chains). This approach eliminates the variability in cellular composition that has complicated studies of primary muscle to date. In a controlled fashion the interactions and contributions of different cell types to the development of normal and genetically dystrophic human muscle can be studied.