Modelling the neurovascular unit and the blood-brain barrier with the unique function of pericytes.
The blood-brain barrier (BBB) is a dynamic cellular complex that is responsible for the maintenance of brain homeostasis. To understand the BBB's key cellular and molecular mechanisms, in vitro models combining endothelial cells and astrocytes can be used to reproduce most of the barrier's 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 BBB's 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.
Université Lille Nord de France, F-59000 Lille, France. elodie.vandenhaute@wanadoo.fr
Elodie
E
Vandenhaute
Lucie
L
Dehouck
Marie-Christine
MC
Boucau
Emmanuel
E
Sevin
Rustem
R
Uzbekov
Meryem
M
Tardivel
Fabien
F
Gosselet
Laurence
L
Fenart
Romeo
R
Cecchelli
Marie-Pierre
MP
Dehouck
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D000941:Antigens / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D001921:Brain / Q000166:cytology; D002417:Cattle; D002478:Cells, Cultured; D062507:Claudin-5; D057167:Claudins / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods; D042783:Endothelial Cells / Q000502:physiology*; D015870:Gene Expression / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D009457:Neuroglia / Q000502:physiology*; D062793:Occludin; D020286:Pericytes / Q000502:physiology*; D010750:Phosphoproteins / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D051381:Rats; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism; D020112:Rhodamine 123 / Q000378:metabolism; D014700:Verapamil / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D062826:Zonula Occludens-1 Protein; D005723:gamma-Glutamyltransferase / Q000378:metabolism; D000099216:Chondroitin Sulfate Proteoglycan 4
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000199:Actins; D000941:Antigens; D062507:Claudin-5; D057167:Claudins; D008565:Membrane Proteins; D062793:Occludin; D010750:Phosphoproteins; D011509:Proteoglycans; D020797:Receptor, Platelet-Derived Growth Factor beta; D020112:Rhodamine 123; D014700:Verapamil; D014746:Vimentin; D062826:Zonula Occludens-1 Protein; D005723:gamma-Glutamyltransferase; D000099216:Chondroitin Sulfate Proteoglycan 4; C491510:Cldn5 protein, rat; C568987:Ocln protein, rat; C569044:Tjp1 protein, rat; C498862:Acta2 protein, rat