Immortalised Human Brain Endothelial Cell Line

Our cells will offer a promising tool for in vitro blood brain barrier models.

Immortalised human brain endothelial cell line.

The objective of this study was to generate an immortal cell line representative of specialized human brain microvascular endothelia forming the blood brain barrier bbb in vivo. Primary and immortalised human brain endothelial cells becs represent two suitable options for studying p gp function in vitro. This review provides a summary of immortalized endothelial cell lines as a guideline for future studies and improvements in the establishment of in vitro blood brain barrier models. Hbmec cells express p gp claudin 1 claudin 3 occludin 55 57 zo 1 54 56 58 ß catenin icam 1 and vcam 1 some of which were also shown under our experimental conditions ve cadherin zo 1 and.

In the screening of four available immortalized human brain capillary endothelial cell lines hbmec proved to be the most suitable and promising cell line for a human in vitro bbb model in terms of barrier tightness and paracellular permeability in a 24 well mono culture system. Our cells displayed both tight junction and efflux transporter functionalities. Immortalized human cerebral microvascular endothelial cell line hcmec d3. Cell differentiation was stimulated by eliminating an immortalization signal.

The neomycin resistant transfected cells overcame proliferative senescence and after a 6 8 wk period of crisis produced immortalization. As a result 36 immortalized endothelial cell lines of human mouse rat porcine and bovine origins were found for the establishment of in vitro blood brain barrier and brain endothelium models. Here we demonstrate that the immortalized human brain endothelial cell line hcmec d3 is a useful alternative to primary brain endothelial cells as a model of the blood brain barrier for studies of central nervous system infection. However the establishment of a reliable human cell line based bbb model has proven to be difficult.

The hcmec d3 bbb cell line has been extensively characterized for brain endothelial phenotype and is a model of human blood brain barrier bbb function. Immortalized human brain capillary endothelial cells can be used as an alternative. A new human brain microvascular endothelial cell line has been established. The limited supply of primary human becs and their instability over passage number make this choice unattractive for medium high throughput studies.

Human capillary and microvascular endothelial cells hcec were transfected with the plasmid psv3 neo coding for the sv40 large t antigen and the neomycin gene. P glycoprotein p gp expression at the blood brain barrier prevents unwanted blood borne toxins and signalling molecules from entering the brain.