RAT HIPPOCAMPAL NEURONS IN DISPERSED CELL-CULTURE
RAT HIPPOCAMPAL NEURONS IN DISPERSED CELL-CULTURE
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DOI：
10.1016/0006-8993(77)90594-7
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发表时间：
1977-01-01
期刊：
影响因子：
2.9
通讯作者：
COWAN, WM
中科院分区：
文献类型：
--
作者：
BANKER, GA;COWAN, WM
建立了一套体外培养的18天或19天大鼠海马神经元培养系统。胰酶消化后，将细胞以低密度电镀于经多聚赖氨酸处理的玻片上，置于浓缩的培养液中。分离的神经元迅速附着在基质上，并启动突起延伸。几乎没有发生再聚集，非神经细胞的数量也很少。除非与组织块共培养，否则神经元只能存活几天；在有海马区外植体存在的情况下，分离的细胞的初始生长得到改善，其在培养中的存活时间延长到约2wk。在这样的培养中，一些细胞形成了一种特征的分支模式，与体内成熟的海马体锥体细胞非常相似。细胞的发育阶段与其在培养过程中的生长有明显的关系。在我们的培养中，在解离前48小时左右已经完成DNA合成并且正在向皮质板迁移的细胞存活得最好。早期有丝分裂后细胞仍在脑室区域内，而已经到达皮质板的细胞生长较差。这个系统不仅可以研究这些细胞的突起形成，而且还可以研究它们在体外形成特定突触的能力和它们表面的生化成分。
An in vitro system was developed for the study of isolated hippocampal neurons from 18 or 19 day rat fetuses. Following trypsinization the cells were plated out at low density on polylysine-treated coverslips in an enriched medium. The isolated neurons rapidly attached to the substrate and initiate process extension. Little reaggregation occurred and the number of non-neuronal cells present was minimal. Unless co-cultured with tissue explants, the neurons survived for only a few days; in the presence of hippocampal explants, the initial growth of the isolated cells was improved and their survival in culture was extended to about 2 wk. Some of the cells in such cultures developed a characteristic branching pattern closely resembling that of maturing hippocampal pyramidal cells in vivo. There was a clear relationship between the stage of the cells'' development and their growth in culture. Cells which had completed DNA synthesis about 48 h before dissociation, and which were in the process of migration to the cortical plate, survived best in our cultures. Early post-mitotic cells which were still within the ventricular zone and cells which had already reached the cortical plate grew poorly. This system may permit the study not only of process formation by these cells, but also of their capacity to form specific synapses in vitro and of the biochemical constituents of their surfaces.