Differential sensitivity of two endothelial cell lines to hydrogen peroxide toxicity: Relevance for in vitro studies of the blood–brain barrier
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Date
2020
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Publisher
MPDI
Abstract
Oxidative stress (OS) has been linked to blood–brain barrier (BBB) dysfunction which in
turn has been implicated in the initiation and propagation of some neurological diseases. In this study,
we profiled, for the first time, two endothelioma cell lines of mouse brain origin, commonly used as
in vitro models of the blood–brain barrier, for their resistance against oxidative stress using viability
measures and glutathione contents as markers. OS was induced by exposing cultured cells to varying
concentrations of hydrogen peroxide and fluorescence microscopy/spectrometry was used to detect
and estimate cellular glutathione contents. A colorimetric viability assay was used to determine
changes in the viability of OS-exposed cells. Both the b.End5 and bEnd.3 cell lines investigated
showed demonstrable content of glutathione with a statistically insignificant difference in glutathione
quantity per unit cell, but with a statistically significant higher capacity for the b.End5 cell line for de
novo glutathione synthesis. Furthermore, the b.End5 cells demonstrated greater oxidant buffering
capacity to higher concentrations of hydrogen peroxide than the bEnd.3 cells. We concluded that
mouse brain endothelial cells, derived from different types of cell lines, differ enormously in their
antioxidant characteristics. We hereby recommend caution in making comparisons across BBB models
utilizing distinctly different cell lines and require further prerequisites to ensure that in vitro BBB
models involving these cell lines are reliable and reproducible.
Description
Keywords
Oxidative stress, Blood–brain barrier, Glutathione, Neurological diseases
Citation
Alamu, O. et al. (2020). Differential sensitivity of two endothelial cell lines to hydrogen peroxide toxicity: Relevance for in vitro studies of the blood–brain barrier. Cells, 9(2), 403. https://doi.org/10.3390/cells9020403