Immortalized cerebellar cells can be induced to display mature neuronal characteristics

Abstract

We have established and characterized a cell line (designated Cb-E1A) that can be induced to display a variety of neuronal characteristics under simple culture conditions. This cell line was generated by retroviral-mediated gene transfer of the adenovirus 12S E1A- immortalizing gene in cerebellar cells isolated from one-week-old rats. Actively dividing cells express the E1A adenovirus protein, and exhibit minimal expression of glial cell markers and low level expression of neuronal cell markers. The immortalized cells can be induced to differentiate by culture in an alternative depolarizing medium or calcium ionophore-containing medium. This caused the expression of neuronal markers to increase rapidly, while glial markers remain unchanged. Under these culture conditions, the Cb-E1A cells also display a variety of other characteristics which suggest that they may provide a good model system for differentiated cerebellar granule neurons. Such neuronal characteristics include a reduction or cessation of mitosis and an increased susceptibility to glutamate toxicity. We think that this novel cell line and differentiation strategy will facilitate future studies of the cellular mechanisms involved in a wide variety of neuronal functions, including development and neurodegenerative disease.