PROPERTIES OF AN IMMORTALIZED VASCULAR ENDOTHELIAL GLIOMA CELL COCULTURE MODEL OF THE BLOOD-BRAIN-BARRIER

In an effort to obtain a useful in vitro model possessing some of the properties of the blood-brain barrier, we have investigated the proper ties and interactions of immortalized cell lines. Immortalised human u mbilical vein endothelial cells (HUVEC-304), in co-culture with rat C6 glioma cells in a two-chambered assembly, form tight junctional compl exes, and develop a permeability barrier having a high transcellular e lectrical resistance. The endothelial cells generate a barrier with gr eatest integrity in the presence of glioma cells, or in the presence o f glioma cell conditioned medium. Under these conditions, the endothel ial cells also display pronounced structural changes which do not occu r in the absence of glioma cells. Morphological alterations include a flattening of cell shape from a cuboidal-type to a squamous-type of ap pearance, and a re-organization of F-actin microfilaments. The integri ty of the barrier can be reversibly disrupted by osmotic shock or by t umor necrosis factor-alpha (TNF-alpha). We interpret these observation s to indicate that co-cultures of immortalized vascular endothelial an d C6 glioma cells provide a model for the investigation of cell-cell i nteractions required for the generation of a barrier having several pr operties of the blood-brain barrier. (C) 1996 Wiley-Liss, Inc.