Chikungunya Virus Propagation Protocol

The CHIKV prototype strain (PS, accession no. AF369024.2) was a generous gift from M.M. Parida (Defense Research Development Establishment [DRDE], Gwalior, India). The Vero (African green monkey kidney epithelial cells) and RAW 264.7 (mouse monocyte/macrophage cells) cells were procured from NCCS (National Center for Cell Science), India. Dulbecco’s modified Eagle’s medium (DMEM) (PAN Biotech, Aidenbach, Germany) supplemented with 10% fetal bovine serum (FBS) (PAN Biotech, Aidenbach, Germany), along with gentamycin and penicillin-streptomycin (PAN Biotech, Aidenbach, Germany) was used to maintain Vero cells. RAW 264.7 cells were maintained in RPMI 1640 (Gibco RPMI 1640 GlutaMAX; Invitrogen, CA) supplemented with 10% FBS (Gibco FBS, Invitrogen, CA), penicillin-streptomycin, and gentamycin. All of the cells were maintained at 37°C with 5% CO₂ in a humidified incubator (Heracell, Thermo Fisher, MA) (51 (link), 52 (link)).