Establishment of permanent astroglial cell lines, able to differentiate in vitro, from transgenic mice carrying the polyoma virus large T gene: An alternative approach to brain cell immortalization

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Abstract
Permanent untransformed cell lines have been established from the cerebral cortex of transgenic mice that carry the polyoma virus large T gene. The immortalized cells described here synthesize laminin and neural cell adhesion molecules and induce primary neurons to develop neuritic processes. As shown by immunofluorescence and immunoblotting assays, they begin to synthesize the glial fibrillary acidic protein (GFAP) after confluence. Double labelling experiments indicated that GFAP expression is reversibly correlated with the arrest of cell division. The present cells also display adrenergic, serotoninergic, and high levels of muscarinic receptors coupled to the phosphatidylinositol signalling pathway. Taken together, our data show that these cell lines constitute homogeneous cell material that has retained the main differentiative, functional, and growth properties of normal astrocytes. Therefore, such clonal untransformed cell lines should be useful for further molecular studies, addressing terminal differentiation of glial cells, glioneuronal interactions, and astroglial expression of receptors for neurotransmitters. Furthermore, we suggest that this approach of cell immortalization by the use of transgenic mice carrying a non‐transforming oncogene might be extended to a variety of cell types.