Induction of blood-brain barrier properties in cultured brain capillary endothelial cells: comparison between primary glial cells and C6 cell line.
The communication between glial cells and brain capillary endothelial cells is crucial for a well-differentiated blood-brain barrier (BBB). It has been suggested that in vitro primary glial cells (GCs) be replaced by the glial C6 cell line to standardise the model further. This study compares directly the structural and functional differentiation of bovine brain capillary endothelial cells (BBCECs) induced by co-culture with rat primary GCs or C6 cells, for the first time. Trans-endothelial electrical resistance (TEER) measurements showed that under no condition were C6 cells able to reproduce TEER values as high as in the presence of GCs. At the same time, permeability of the BBCECs to both radioactive sucrose and FITC-inulin was 2.5-fold higher when cells were co-cultured with C6 than with GCs. Furthermore, immunocytochemistry studies showed different cell morphology and less developed tight junction pattern of BBCECs co-cultured with C6 toward GCs. Additionally, studies on P-glycoprotein (P-gp) showed much lower P-gp presence and activity in BBCECs co-cultured with C6 than GCs. Both VEGF mRNA expression and protein content were dramatically increased when compared with GCs, suggesting that VEGF could be one of the factors responsible for higher permeability of BBB. Our results clearly indicate that, in the presence of the glial C6 cell line, BBCECs did not differentiate as well as in the co-culture with primary GCs at both structural and functional levels.
Boveri
Monica
M
ECVAM, Institute for Health and Consumer Protection, Joint Research Centre, European Commission, Ispra (VA), Italy.
Berezowski
Vincent
V
Price
Anna
A
Slupek
Stephanie
S
Lenfant
Anne-Marie
AM
Benaud
Christelle
C
Hartung
Thomas
T
Cecchelli
Romeo
R
Prieto
Pilar
P
Dehouck
Marie-Pierre
MP
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001812:Blood-Brain Barrier / Q000166:cytology / Q000378:metabolism*; D001921:Brain / Q000098:blood supply / Q000502:physiology*; D002417:Cattle; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism; D007444:Inulin / Q000493:pharmacokinetics; D008564:Membrane Potentials / Q000502:physiology; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013395:Sucrose / Q000493:pharmacokinetics; D015854:Up-Regulation / Q000502:physiology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D013395:Sucrose; D007444:Inulin