Development and characterization of clonal cell lines derived from septal cholinergic neurons

Studies employing primary cells to determine the molecular basis of neuronal development and selective synaptogenesis in the central nervous system are limited by cellular heterogeneity. Clonal hybrid cell lines derived from a particular region of brain, which express differentiated characteristics typical of the cells of origin, offer a potentially powerful alternative approach. We previously demonstrated the feasibility of deriving such cell lines from septal cholinergic cells. We now delineate the methods employed, and describe the development of additional cholinergic cell lines expressing neuronal and cholinergic features from later developmental stages. One cell line has been studied in detail and found to form neurites, express choline acetyltransferase (ChAT) and neurofilament protein (NFP), and display typical neuronal ultrastructural characteristics, including puncta adherens, neuritic varicosities, vesicles, and growth cones.