Pegolo G, Askanas V, Engel W K
Department of Neurology, University of Southern California School of Medicine, Hospital of the Good Samaritan, Los Angeles 90017.
Int J Dev Neurosci. 1990;8(3):299-308. doi: 10.1016/0736-5748(90)90036-2.
Primary cultures of muscle cells derived from a biopsied adult human skeletal muscle were grown up to 6 weeks in a hormonally/chemically enriched serum-free medium. The expression of muscle-specific isozymes of creatine kinase, glycogen phosphorylase, and phosphoglycerate mutase, indicative of muscle cell maturation, was studied after 1, 4 and 6 weeks of growth. The maturation of muscle fibers cultured in serum-free medium was comparable to that achieved by muscle fibers cultured in medium containing 10% serum and supplemented with growth factors (insulin, epidermal growth factor, and fibroblastic growth factor) and was greater than that achieved in medium containing 10% serum only. Our study demonstrates that adult human muscle can be cultured aneurally for a long period of time in a serum-free medium, and that it can achieve a high degree of maturation. This study provides an important basis for investigations related to: (1) assessment of the influence of individual components of the medium on human muscle maturation in culture; (2) studies of regulation of abnormal gene expression in diseased human muscle cultured in serum-fre medium.
取自活检的成人人类骨骼肌的肌肉细胞原代培养物,在富含激素/化学物质的无血清培养基中培养长达6周。在生长1周、4周和6周后,研究了肌酸激酶、糖原磷酸化酶和磷酸甘油酸变位酶等肌肉特异性同工酶的表达,这些同工酶可指示肌肉细胞的成熟情况。在无血清培养基中培养的肌纤维成熟情况,与在含有10%血清并添加生长因子(胰岛素、表皮生长因子和成纤维细胞生长因子)的培养基中培养的肌纤维相当,且优于仅含有10%血清的培养基中培养的肌纤维。我们的研究表明,成人人类肌肉可以在无血清培养基中长时间无神经培养,并能达到高度成熟。本研究为以下相关研究提供了重要依据:(1)评估培养基中各个成分对培养的人类肌肉成熟的影响;(2)研究在无血清培养基中培养的患病人类肌肉中异常基因表达的调控。