Immortalization of Spinal Cord Motor Neurons by Fusion of Primary Neurons with Neuroblastoma Cell Lines

Abstract

The study of motor neurons is a technically difficult area in neurobiology because of limitations in isolation and culture. Among the many approaches used for isolating and culturing primary motor neurons, few have produced enough cells for biochemical and molecular studies. To overcome this problem, we have used somatic cell hybridization methods to generate hybrid neural cells that have traits typical of motor neurons. Isolated embryonic mouse spinal cord motor neurons were fused to an aminopterin-sensitive and neomycin-resistant mouse neuroblastoma cell line to produce several hybrid neuron cell lines. Those cell lines expressing high levels of choline acetyltransferase activity were selected and cloned. The hybrid nature of the cloned cells was confirmed by analysis of glucose phosphate isomerase allozymes and karyotyping. Availability of these embryonic clonal hybrid cells will make possible molecular, physiologic, and biochemical studies to define the biological properties of spinal motor neurons.