Human Cerebral Organoids Reveal New Understanding of Coronavirus OC43 Infection adn Potential Treatments
A new study published in the Journal of Biomedical Science details the effects of Human Coronavirus OC43 (HCoV-OC43) infection on human cerebral organoids, offering novel insights into the pathogenesis of the virus and potential therapeutic interventions. Researchers utilized these 3D in vitro models of the human brain too investigate the virus’s impact on the blood-brain barrier (BBB).
Methods Employed to Model Infection and Assess BBB Integrity
Human brain microvascular endothelial cells (HBMECs) were cultured on transwell inserts to model the BBB. The integrity of the BBB was assessed using trans-epithelial electrical resistance (TEER) measurements.The TEER probe was washed with 75% (v/v) ethanol,air-dried,and equilibrated in endothelial medium for 15 minutes. Measurements were taken at multiple points across each Transwell insert, and TEER values were recorded daily.
BBB permeability was further evaluated by measuring the penetration rate of FITC-Dextran,following a protocol established by Reka Natarajan’s group [38]. Specifically, DMEM/F12 (without phenol red) containing 100 µg/mL of FITC-Dextran was applied to the apical side for 30 minutes, and fluorescence intensity was measured using a microplate reader at excitation/emission wavelengths of 494/521 nm. Permeability was calculated as (apical fluorescence intensity/basal fluorescence intensity) × 100%.
immunostaining was performed on HCoV-OC43 infected BBB cells using antibodies against GFAP (CST 80788T, 1:100), claudin 5 (ThermoFisher 35-2500, 1:100), and CD146 (abcam ab75769, 1:100).Samples were visualized using a leica TCS SP8 laser scanning confocal microscope.
Statistical Analysis
Data are presented as mean ± SD, with individual data points shown. An unpaired t-test was used for comparisons between two groups, and one-way ANOVA for three or more groups.Statistical significance was set at *p* < 0.05, with analyses performed using GraphPad Prism 9.5.
