Cultivation of BHK-21 and CHIKV Replicon Cells

BHK-21 cells (fibroblasts derived from Syrian golden hamster kidney; ATCC CCL-10), purchased from The Global Bioresource Center (ATCC), were maintained in Dulbecco’s modified Eagle’s medium (DMEM, SIGMA-ALDRICH) supplemented with 100U/mL of penicillin (HYCLONE LABORATORIES), 100 mg/mL of streptomycin (HYCLONE LABORATORIES), 1% dilution of stock of non-essential amino acids (Hyclone Laboratories) and 1% of fetal bovine serum (FBS, HYCLONEN LABORATOIRES) in a humidified 5% CO₂ incubator at 37 °C. Subgenomic replicon (SGR) harboring cell lines (BHK-CHIKV-NCT)^{37 (link)} were maintained under the same conditions of BHK-21 cells (ATCC CCL-10), except for the addition of G418 (SIGMA-ALDRICH) at 5 mg/mL.