Characterisation of an in vitro blood-brain barrier model based on primary porcine capillary endothelial cells in monoculture or co-culture with primary rat or porcine astrocytes and pericytes.

In vitro blood-brain barrier (BBB) models based on primary brain capillary endothelial cells (BCECs) in monoculture or in co-culture with primary astrocytes and pericytes are often applied for studying physiology of the BBB. Primary BCECs retain many morphological and biochemical properties similar to in vivo such as efflux transporters, tight junction proteins, and high transendothelial electric resistance (TEER). Primary BCECs are isolated from a variety of mammals such as rats, mice, cattle and pigs. Often bovine and porcine BCECs are cultured in monoculture or in co-culture with rat astrocytes obtained from neonatal rats which have been shown to strengthen the barrier properties of the BCECs.
In this study, brain endothelial cells (PBECs), astrocytes and pericytes are isolated from pig brains donated by the local abattoir. The brains are from 6 month old domestic pigs. The availability and high yield of cells per isolation makes the pigs a favourable source for BCECs, astrocytes and pericytes.
The aim of this study was to determine which type of culture method would result in the most optimal porcine in vitro BBB model. The effect of co-culture of PBECs with rat astrocytes and/or pericytes was compared with co-culture of PBECs with porcine astrocytes and/or pericytes. These results were also compared to results obtained from monoculture of PBECs. The effect of both contact and non-contact co-culture with porcine or rat astrocytes and pericytes was investigated. The different culture methods were evaluated on the basis of TEER measurements, PCR and immunocytochemical staining of BCEC specific proteins.