A Protocol Using Retrovirally Introduced Multiple Oncogenes for Producing Neuron-like Cell Lines from the Murine Central Nervous System

Abstract

A useful approach to molecular biologic studies of complex mammalian systems is the judicious use of clonal cell lines. For studies on neuronal populations of the central nervous system, useful cell lines are lacking. A general protocol that produces clonal lines by retroviral infection in vivo followed by in vitro infection with an additional oncogene-containing virus is detailed. The lines produced express several neuronal markers but not glial fibrillary acidic protein. The lines are phenotypically stable, freezable, and transfectable by standard methods. Retroviral combinations that have produced stable lines using this protocol are large T with v-src and v-myc, and large T with v-ras. Other oncogenic combinations may produce different, useful phenotypes for studies on the molecular development and function of CNS neurons.