The blood-brain barrier (BBB) is a dynamic cellular complex that is responsible for the maintenance of brain homeostasis. To understand the BBBs key cellular and molecular mechanisms, in vitro models combining endothelial cells and astrocytes can be used to reproduce most of the barriers in vivo features (low paracellular permeability and the expression of specific transporters). However, these models lack pericytes - a poorly characterized cell type which appears to be of crucial importance to understand BBBs function in healthy and diseased states. The present study sought to identify and characterize this cell population - which lacks a specific marker - by comparing its phenotype with that of vascular smooth muscle cells. Even if pericytes and smooth muscle cells shared many markers in vitro, our results showed that they could be distinguished by their different P-glycoprotein expression and γ-glutamyltranspeptidase activity. Two different three-cell-type culture models were described, including pericytes to mimic the neurovascular unit. In the first model, endothelial cells were cultured alone on a filter, away from glial cells and pericytes, allowing endothelial cell phenotype characterization. In the second model, glial cells were at the bottom of the well while pericytes and endothelial cells were cultured together in the filter: close interactions were observed in peg-and-socket contacts. In both models low paracellular permeability and P-glycoprotein functionality were demonstrated. These models are likely to be useful tools for understanding the pericytes role in BBB physiology and could be of value in investigating the pericytes influence on BBB in diseased states.
Keywords: Blood-brain barrier, endothelial cells, in vitro models, neurovascular unit, pericytes, three-cell-type model, pinocytic activity, efflux pumps, homeostasis, Alzheimer's disease, neurodegeneration, gamma-glutamyltranspeptidase, Non-Contact Model, manipulation