]. Briefly, blood vessels and meninges were removed from the foetal brain tissue . Thereafter, the tissue was minced, treated with 0.2 mg/ml DNase I and 0.25% trypsin for 30 min before being passed through a 70-µm cell strainer . The flow-through was plated in petri dishes for adherent cells at a final concentration of 6–8 × 10cells/petri in MEM supplemented with 10% FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 0.
] with some modifications. Briefly, cells were seeded in clear bottom, black 96-well plates at 2 × 10cells/well and cultured for 24 h before virus infection. Cells were then incubated with SARS-CoV-2 isolates at various MOI for 24 h before being washed thoroughly with PBS to remove unbound virus. Cells were then fixed with 4% formaldehyde for 1 h after 24 h , 2 days, 4 days, or 6 days of infection. Cells were subsequently permeabilised with 0.
]. Whole-well imaging at 5 × magnification was acquired using an Opera Phenix and fluorescent areas calculated using Harmony v4.9 software .CNS cells seeded either on inserts model, described later), E-plates or 96-well plates were infected with SARS-CoV-2 isolates at two MOI for up to 6 days. MTS, BBB permeability and glutamate assays were performed after 2, 4 and 6 days of infection. For the xCELLigence assay, the impedance was continuously recorded for 6 days after infection.