Human astrocytes/astrocyte-conditioned medium and shear stress enhance the barrier properties of human brain microvascular endothelial cells.
The blood-brain barrier (BBB) is a structural and functional barrier that regulates the passage of molecules into and out of the brain to maintain the neural microenvironment. We have previously developed the in vitro BBB model with human brain microvascular endothelial cells (HBMEC). However, in vivo HBMEC are shown to interact with astrocytes and also exposed to shear stress through blood flow. In an attempt to develop the BBB model to mimic the in vivo condition we constructed the flow-based in vitro BBB model using HBMEC and human fetal astrocytes (HFA). We also examined the effect of astrocyte-conditioned medium (ACM) in lieu of HFA to study the role of secreted factor(s) on the BBB properties. The tightness of HBMEC monolayer was assessed by the permeability of dextran and propidium iodide as well as by measuring the transendothelial electrical resistance (TEER). We showed that the HBMEC permeability was reduced and TEER was increased by non-contact, co-cultivation with HFA and ACM. The exposure of HBMEC to shear stress also exhibited decreased permeability. Moreover, HFA/ACM and shear flow exhibited additive effect of decreasing the permeability of HBMEC monolayer. In addition, we showed that the HBMEC expression of ZO-1 (tight junction protein) was increased by co-cultivation with ACM and in response to shear stress. These findings suggest that the non-contact co-cultivation with HFA helps maintain the barrier properties of HBMEC by secreting factor(s) into the medium. Our in vitro flow model system with the cells of human origin should be useful for studying the interactions between endothelial cells, glial cells, and secreted factor(s) as well as the role of shear stress in the barrier property of HBMEC.
Siddharthan
Venkatraman
V
Division of Pediatric Infectious Diseases, Johns Hopkins University, School of Medicine, Baltimore, MD 21287, USA.
Kim
Yuri V
YV
Liu
Suyi
S
Kim
Kwang Sik
KS
D001253:Astrocytes / Q000502:physiology*; D001685:Biological Factors / Q000378:metabolism; D001812:Blood-Brain Barrier / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D001921:Brain / Q000098:blood supply / Q000166:cytology / Q000502:physiology*; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003911:Dextrans / Q000493:pharmacokinetics; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007202:Indicators and Reagents / Q000493:pharmacokinetics*; D011419:Propidium / Q000493:pharmacokinetics; D033081:Shear Strength; D013314:Stress, Mechanical
D001685:Biological Factors; D017077:Culture Media, Conditioned; D003911:Dextrans; D007202:Indicators and Reagents; D011419:Propidium