Saneto R P, de Vellis J
Proc Natl Acad Sci U S A. 1985 May;82(10):3509-13. doi: 10.1073/pnas.82.10.3509.
A serumless, chemically defined medium has been developed for the culture of oligodendrocytes isolated from primary neonatal rat cerebral cultures. Combined together, insulin, transferrin, and fibroblast growth factor synergistically induced an essentially homogenous population (95-98%) of cells expressing glycerol-3-phosphate dehydrogenase (EC 1.1.1.8) activity to undergo cell division. Proliferating cels were characterized by several criteria: (i) ultrastructural analysis by transmission electron microscopy identified the cell type as an oligodendrocyte; (ii) biochemical assays showed expression of three oligodendrocyte biochemical markers, induction of both glycerol phosphate dehydrogenase and lactate dehydrogenase (EC 1.1.1.27), and presence of 2',3'-cyclic nucleotide 3'-phosphodiesterase (EC 3.1.4.37); and (iii) immunocytochemical staining showed cultures to be 95-98% positive for glycerol phosphate dehydrogenase, 90% for myelin basic protein, 60-70% for galactocerebroside, and 70% for A2B5. Few cells (less than 5%) stained positive for glial fibrillary acidic protein, and none were detected positive for fibronectin.
已开发出一种无血清、化学成分明确的培养基，用于培养从新生大鼠原代脑培养物中分离出的少突胶质细胞。胰岛素、转铁蛋白和成纤维细胞生长因子共同作用，协同诱导表达甘油-3-磷酸脱氢酶（EC 1.1.1.8）活性的细胞群体（95-98%）基本均匀地进行细胞分裂。增殖细胞通过以下几个标准进行表征：（i）通过透射电子显微镜进行的超微结构分析确定细胞类型为少突胶质细胞；（ii）生化分析显示三种少突胶质细胞生化标志物的表达、甘油磷酸脱氢酶和乳酸脱氢酶（EC 1.1.1.27）的诱导以及2',3'-环核苷酸3'-磷酸二酯酶（EC 3.1.4.37）的存在；（iii）免疫细胞化学染色显示培养物中甘油磷酸脱氢酶阳性率为95-98%，髓鞘碱性蛋白为90%，半乳糖脑苷脂为60-70%，A2B5为70%。很少有细胞（少于5%）对胶质纤维酸性蛋白染色呈阳性，未检测到对纤连蛋白呈阳性的细胞。
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