ISOLATION AND CHARACTERIZATION OF CONDITIONALLY IMMORTALIZED ASTROCYTE CELL-LINES DERIVED FROM ADULT HUMAN SPINAL-CORD

As an approach to develop both oligodendrocytic and astrocytic cell li nes from adult human spinal cord, a cellular preparation of highly enr iched oligodendrocytes and their precursors was infected with a replic ation-deficient retrovirus containing DNA sequences encoding the tempe rature-sensitive mutant of SV40 large T antigen. Six immortal cell lin es were obtained. At both permissive (33 degrees C) and non-permissive (38.5 degrees C) temperatures, all cell lines were positive for vimen tin, two demonstrated glial fibrillary acidic protein (GFAP) immunorea ctivity, and none expressed oligodendrocyte or microglial markers. The 2 GFAP-positive cell lines [human spinal cord (HSC)2 and HSC6] were f urther characterized. Karyotype analysis revealed that both HSC2 and H SC6 cells showed gain of chromosomal material and structural chromosom al abnormalities. However, at non-permissive temperature both cell lin es were indistinguishable from primary human astrocytes by a number of criteria. These properties included glutamine synthetase activity, Na +-dependent glutamate uptake, K+ flux, purine-evoked Ca2+ mobilization and entry, and the ability to support neurite outgrowth from embryoni c rat retinal explants. The HSC2 and HSC6 cell lines may prove to be v aluable models for studying the physiological properties of adult huma n astrocytes. (C) 1994 Wiley-Liss, Inc.