Immortalized Human Cerebromicrovascular Endothelial Cells

Primary cultures of cerebromicrovascular endothelial cells (CEC) derived from rat, bovine, porcine, mouse and human can rapidly lose key phenotypic markers of the blood-brain barrier (BBB) and undergo cellular senescence after a limited number of divisions in vitro. Furthermore, expression of BBB markers varies considerably among BCEC obtained from different species. These issues, compounded with the problems associated with very limited availability of human brain biopsies, small initial yield of cells and short proliferative life span of human cells, greatly restrict the utility of primary human BCEC as a reliable in vitro BBB model. Therefore, Muruganandam et al. (1997) developed an immortalized human cerebromicrovascular endothelial cell line as an in vitro model of the human blood-brain barrier. 

The permeability coefficient Pe [cm/min] for the endothelial monolayer is derived by dividing PSe by surface area (0.83 cm2) of the membranes. 

Permeability coefficients for paracellular diffusion of 3H-sucrose and 14C-inulin across monolayers of HCEC and immortalized SV-HCEC and transendothe-lial resistances have been compared to human lung microvascular endothelial cells to evaluate the BBB phenotype of the cerebromicrovascular endothelial cell cultures (Muruganandam et al. 1997). 

MODIFICATIONS OF THE METHOD Rat brain capillary endothelial cells have been immortalized by the entire E1A region of adenovirus 2 by Roux et al. (1994) which was used for a number of cell types without oncogenic transformation. Bovine umbilical vein endothelial cells have been immortalized by transfection with an expression vector containing 

Cajero-Juarez M, Avila B, Ochoa A et al. (2002) Immortalization of bovine umbilical vein endothelial cells a model for the study of vascular endothelium. Eur J Cell Biol 81 1-8 Gerhart DZ, Broderius MA, Drewes LR (1988) cultured human and canine endothelial cells from brain microvessels. Brain Res BuU 21 785-793 

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Muruganandam, A., L. M. Herx, R. Monette, J. P. Durkin, and D. B. Sta-nimirovic. 1997. Development of immortalized human cerebromicrovascular endothelial cell line as an in vitro model of the human blood-brain barrier. Faseb J 11(13) 1187—97. 

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