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Real‐time acquisition of transendothelial electrical resistance in an all‐human, in vitro , 3‐dimensional, blood‐brain barrier model exemplifies tight‐junction integrity
Author(s) -
Maherally Zaynah,
Fillmore Helen L.,
Tan Sim Ling,
Tan Suk Fei,
Jassam Samah A.,
Quack Friederike I.,
Hatherell Kathryn E.,
Pilkington Geoffrey J.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.201700162r
Subject(s) - occludin , tight junction , blood–brain barrier , microbiology and biotechnology , laminin , in vitro , fibronectin , human brain , endothelial stem cell , biology , chemistry , biophysics , extracellular matrix , central nervous system , neuroscience , biochemistry
The blood‐brain barrier (BBB) consists of endothelial cells, astrocytes, and pericytes embedded in basal lamina (BL). Most in vitro models use nonhuman, monolayer cultures for therapeutic‐delivery studies, relying on transendothelial electrical resistance (TEER) measurements without other tight‐junction (TJ) formation parameters. We aimed to develop reliable, reproducible, in vitro 3‐dimensional (3D) models incorporating relevant human, in vivo cell types and BL proteins. The 3D BBB models were constructed with human brain endothelial cells, human astrocytes, and human brain pericytes in mono‐, co‐, and tricultures. TEER was measured in 3D models using a volt/ohmmeter and cellZscope. Influence of BL proteins—laminin, fibronectin, collagen type IV, agrin, and perlecan—on adhesion and TEER was assessed using an electric cell‐substrate impedance‐sensing system. TJ protein expression was assessed by Western blotting (WB) and immunocytochemistry (ICC). Perlecan (10 μg/ml) evoked unreportedly high, in vitro TEER values (1200 Ω) and the strongest adhesion. Coculturing endothelial cells with astrocytes yielded the greatest resistance over time. ICC and WB results correlated with resistance levels, with evidence of prominent occludin expression in cocultures. BL proteins exerted differential effects on TEER, whereas astrocytes in contact yielded higher TEER values and TJ expression.—Maherally, Z., Fillmore, H. L., Tan, S. L., Tan, S. F., Jassam, S. A., Quack, F. I., Hatherell, K. E., Pilkington, G. J. Real‐time acquisition of transendothelial electrical resistance in an all‐human, in vitro , 3‐dimensional, blood‐brain barrier model exemplifies tight‐junction integrity. FASEB J. 32, 168‐182 (2018). www.fasebj.org