Cyclic morphological changes of glial cells in long-term cultures of rat brain

Brain cells from embryonic rats were dissociated with trypsin, cultivated under constant conditions in Falcon flasks, and studied for periods of one year or more. Antisera against glial fibrillary acidic protein (GFA) and myelin basic protein (MBP) were used to identify glial cell types. For scanning electron microscopical (SEM) observation an embedding method in resin was developed that allows good preservation of the fine ultrastructural features of the cultivated cells and precise characterization of the cell types. Under our culture conditions, after four subcultures and 8-10 weeks of cultivation, the following cell types can be distinguished. (a) Flat epitheloid cells. From an immunocytological point of view these cells form a heterogeneous population composed of GFA- and MBP-positive and negative cells. They are the precursors of the following cell types. (b) Astroglial cells. SEM observations show a characteristic network of radially orientated prolongations. 92% of these cells are GFA-positive. (c) Oligodendroglial cells with characteristic dichotomously dividing branches. Secondary and tertiary branches end in flat amoeboid prolongations. These cells are MBP-positive. After approximately six weeks the most prominent cells are the flat epitheloid cells. The astroglial cells originate continuously from the epitheloid cells during the whole cultivation time. The formation of oligodendroglial cells, on the other hand, takes place at relatively precise intervals of time (approximately every 20-30 days) over the entire cultivation period (of more than one year).