A new blood–brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes

Abstract

Blood-brain barrier (BBB) characteristics are induced and maintained by cross-talk between brain microvessel endothelial cells and neighbouring elements of the neurovascular unit. While pericytes are the cells situated closest to brain endothelial cells morphologically and share a common basement membrane, they have not been used in co-culture BBB models for testing drug permeability. We have developed and characterized a new syngeneic BBB model using primary cultures of the three main cell types of cerebral microvessels. The co-culture of endothelial cells, pericytes and astrocytes mimick the anatomical situation in vivo. In the presence of both pericytes and astrocytes rat brain endothelial cells expressed enhanced levels of tight junction (TJ) proteins occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. Further morphological evidence of the presence of interendothelial TJs was provided by electron microscopy. The transendothelial electrical resistance (TEER) of brain endothelial monolayers in triple co-culture, indicating the tightness of TJs reached 400Omegacm(2) on average, while the endothelial permeability coefficients (P(e)) for fluorescein was in the range of 3x10(-6)cm/s. Brain endothelial cells in the new model expressed glucose transporter-1, efflux transporters P-glycoprotein and multidrug resistance protein-1, and showed a polarized transport of rhodamine 123, a ligand for P-glycoprotein. To further characterize the model, drug permeability assays were performed using a set of 19 compounds with known in vivo BBB permeability. Good correlation (R(2)=0.89) was found between in vitroP(e) values obtained from measurements on the BBB model and in vivo BBB permeability data. The new BBB model, which is the first model to incorporate pericytes in a triple co-culture setting, can be a useful tool for research on BBB physiology and pathology and to test candidate compounds for centrally acting drugs.

Retinoic acid and hydrocortisone strengthen the barrier function of human RPMI 2650 cells, a model for nasal epithelial permeability

“…This finding is in agreement with data on the favourable effect of hydrocortisone on epithelial cells described previously (Wu et al 1986;Van Scott et al 1988). Hydrocortisone is a potent inducer of the formation of barrier properties in cultured endothelial cells (Perrière et al 2007;Nakagawa et al 2009), and the increase in the cell index of RPMI 2650 layers may be also related to the formation of a tighter barrier. Cholera toxin, which acts through cAMP as a second messenger, improves cell growth and inhibits the differentiation-inducing activity of serum (Lechner 1984;Wu et al 1986).…”

Transfer of Opiorphin Through a Blood-Brain Barrier Culture Model

“…The resistance data and P app ARCMED2032_proof ■ 30-6-2015 20-16-19 values for passive hydrophilic permeability markers fluorescein and albumin were in accordance with our previous data and indicated a tight barrier (16,17). Instead of radiolabeling we used the native form of the peptide and a sensitive method, LC-MS, to detect opiorphin.…”

“…Cell isolation and the preparation of the co-culture BBB model was performed as previously described (16). Brain endothelial cells and pericytes were seeded on the opposite surfaces of collagen IV and fibronectin coated Costar Transwell polycarbonate inserts (12 mm diameter, 0.4 mm pore size; Corning, Corning, NY) and kept in co-culture with glial cells to reach good barrier properties for the permeability measurements ( Figure 1A).…”

A versatile lab-on-a-chip tool for modeling biological barriers

“…We established a syngeneic rat BBB model based on the co-culture of primary brain endothelial cells with pericytes and glial cells mimicking the in vivo anatomical position of the cells. This triple co-culture BBB model displays barrier properties and in drug permeability assays shows good correlation with in vivo BBB permeability data [14,15].…”