IN-VITRO NEURONAL AND GLIAL PRODUCTION AND DIFFERENTIATION OF HUMAN CENTRAL NEUROCYTOMA CELLS

Human central neurocytoma cells were cultured and characterized immuno phenotypically and electrophysiologically to clarify their development al potential, We conducted systematic in vitro studies utilizing fresh tissues from three patients, Initially small homogenous cell clusters settled down onto the bottom of the culture flasks, and, after 2 week s from plating, mature neuron-like cells developed from these clusters and expressed neurofilament proteins (NF: specific neuronal markers), On the other hand, approximately 80% of small round cell clusters and flat glial-like cells from which these clusters developed were positi vely stained for glial fibrillary acidic protein (GFAP: a specific gli al marker), Furthermore these neuronal and glial cells showed distinct morphology, and dual-label, indirect immunohistochemistry for GFAP an d NF-200 kD disclosed that the two antigens were not found co-localize d in the same cells, In single-cell clonal analysis, neuronal, glial, and mixed neuronal and glial clones were generated, Electrophysiologic ally, the cells of neuronal morphology possessed sodium channels, and also L-type calcium channels in whole-cell voltage clamp, The sodium c hannels were of a characteristic neuronal phenotype which appears in n eurons, These findings suggest that small round human central neurocyt oma cells exhibit both neuronal and glial differentiations and have th e properties reminiscent of precursor cells derived from subventricula r matrix; thus, these cultured cells may be a potential source for inv estigations of human CNS neuronal and glial development and differenti ation. (C) 1998 Wiley-Liss, Inc.