Jeserich G, Rauen T
Department of Zoophysiology, University of Osnabrück, Federal Republic of Germany.
Glia. 1990;3(1):65-74. doi: 10.1002/glia.440030109.
Oligodendrocytes were isolated from the white matter of young trout by Percoll density centrifugation of enzymatically dissociated tissue and cultured on poly-D-lysine-coated petri dishes. Using antisera recognizing myelin-specific compounds of fish CNS (36K, IP2) up to 72% of the isolated cells could be identified as oligodendrocytes with an average yield of 4 x 10(6) cells per gram of wet tissue. Taken in culture, the cells rapidly regenerated their processes and soon acquired a morphology closely resembling mammalian oligodendrocytes in vitro. On the other hand, in terms of phenotypic expression, interesting parallels were revealed with the known in vitro behavior of Schwann cells: Galactocerebroside, which in mammalian oligodendrocytes is persistently expressed over longer periods of time in vitro, rapidly disappeared from the surface of cultured trout oligodendrocytes. In contrast, the fish CNS myelin glycoprotein IP2, which like IP1 is immunologically related to the major myelin product of Schwann cells, P0, was continuously expressed over several weeks in culture. Two other myelin protein constituents, 36K and IP1, transiently declined in vitro, but later on fully reappeared in the glial cells of trout. The present cell culture system offers an experimental model for studying in vitro the factors underlying oligodendroglial regeneration and remyelination in the fish CNS.
通过对酶解组织进行Percoll密度离心,从幼年鳟鱼的白质中分离出少突胶质细胞,并将其培养在聚-D-赖氨酸包被的培养皿上。使用识别鱼类中枢神经系统髓鞘特异性化合物(36K、IP2)的抗血清,高达72%的分离细胞可被鉴定为少突胶质细胞,平均每克湿组织的产量为4×10⁶个细胞。在培养过程中,这些细胞迅速再生其突起,并很快获得了与体外培养的哺乳动物少突胶质细胞非常相似的形态。另一方面,在表型表达方面,发现与雪旺细胞已知的体外行为存在有趣的相似之处:半乳糖脑苷脂在哺乳动物少突胶质细胞体外长时间持续表达,但在培养的鳟鱼少突胶质细胞表面迅速消失。相反,鱼类中枢神经系统髓鞘糖蛋白IP2,与IP1一样,在免疫上与雪旺细胞的主要髓鞘产物P0相关,在培养的几周内持续表达。另外两种髓鞘蛋白成分3K和IP1在体外短暂下降,但后来在鳟鱼的神经胶质细胞中完全重新出现。目前的细胞培养系统为体外研究鱼类中枢神经系统少突胶质细胞再生和髓鞘再生的潜在因素提供了一个实验模型。