Abstract
In vitro neurovirological studies of viral infectivity or viral gene expression may be confounded by the multiple neural cell type and/or fibroblast contamination present in early passage culture prepared from dissociated human central nervous system (CNS) tissue. We have development highly enriched astrocyte cultures for neurovirological study by culturing in a serum-free defined medium, B16, supplemented with basic fibroblast growth factor (FGF-2). subculture in this medium selects against fibroblast proliferation and favors sustained proliferation of a highly enriched gliaf fibrillary acidic protein (GFAP)-positive cell population. These astrocytes suppory productive replication of cytomegalovirus (CMV) and transient expression of transfected CMV and human imunodeficiency virus type 1 (HIV-1) viral promoters. By comparison. CNS cultures developed in standard serum-containing medium initially contain predominantly astrcytes, but show increasing contamination with fibroblasts with sequential passage. These cultures support CMV viral synthesis in both fibroblasts and astrocytes, cell types distinguishable only by immunostaining for cell specifica antigen. CMV or HIV-1 promoter activities, quantitated by transient gene expression assays, are distinctly lower in CNS cultures maintained in serum-containing medium
