Fast type I interferon response protects astrocytes from flavivirus infection and virus-induced cytopathic effects

Lindqvist, Richard; Mundt, Filip; Gilthorpe, Jonathan D.; Wölfel, Silke; Gekara, Nelson O.; Kröger, Andrea; Överby, Anna K.

The mice were sacrificed between postnatal day 1 and 4 for astrocyte isolation. Cerebral cortices were isolated, and cells were seeded in poly-D-lysine (Sigma) coated T75 tissue culture flasks as previously described [40] . Monolayers of astrocytes were shaken at 200 rpm for 1 h to remove microglia and oligodendrocyte precursors before seeding for experiments. Primary cortical neurons were derived from cerebral cortices of embryonic day 17 mice. The cortices were isolated, and cells were seeded in Dulbecco's modified Eagle's medium (DMEM, Sigma D5648-10L) containing 10 % heat-inactivated foetal bovine serum (FBS, Gibco) and 0.1 U/mL penicillin and 0.1 μg/mL streptomycin (Gibco) in poly-D-lysine (Sigma) coated 96-well plates as previously described [41] . After 3 h, the medium was replaced with Neurobasal medium (Gibco) containing B27 (Gibco), 0.1 U/mL penicillin, 0.1 μg/mL streptomycin, and 2 mM L-glutamine (Gibco). The neurons were infected at day 7 post seeding.