Blood-brain barrier in vitro model utilizing immortalized cell lines

Abstract

The interest in using blood-brain barrier (BBB) in vitro culture assays in early safety assessment of drugs is growing [1]. This study evaluated the feasibility of using commercially available immortalized cell lines instead of primary cell lines in building in vitro models for BBB. Immortalized mouse endothelial cells from yolk sac and rat astrocytes from 1-day old rat purchased from American Type Culture Collections (ATCC) were used in the assessment to build the BBB co-culture in vitro model. Acellular extracellular matrix (aECM) derived from a confluent layer of astrocytes was introduced in the experiment providing a novel biomimetic basement membrane scaffold for endothelial cells. Experimental data shown in this paper were based on experiments with at least three set of replications. The BBB in vitro co-culture model with aECM is promising in terms of demonstrating higher daily transendothelial electrical resistance (TEER) values, and better inhibition of solute transport of different compounds than endothelial cells cultured alone as well as traditional co-cultured models. Immunofluorescence study and Western Blot analysis also verified higher expression level of occludin, one of the tight junction proteins in the co-cultured model with aECM than any other sample setups. Our novel approach using immortalized cell lines with the addition of aECM was testified to be a cost and labor effective, feasible and repeatable alternative for BBB in vitro model building in drug delivery studies. © 2009 Springer Berlin Heidelberg.