Atcc ccl 10

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.

African green monkey kidney cells [Vero; ATCC CCL-81 from American Type Culture Collection (ATCC), Rockville, MD] and BHK-21 cells (ATCC CCL-10 from ATCC) were cultured at 37°C in 5% CO₂ in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco), 2-mM L-glutamine (Life Technologies) and antibiotics (100-U ml⁻¹ penicillin and 100-μg ml⁻¹ streptomycin; Life Technologies). The KOS strain of HSV-1 was used as the wild-type strain. The K26GFP HSV-1 recombinant virus (gift from Dr. Prashant Desai), which carries a GFP tag on the capsid protein VP16, was used in fluorescence studies. All viruses were amplified on Vero cells, and titers were determined on Vero cells by plaque assay. Viral plaque assays were carried out as follows: Viral stocks were serially diluted in DMEM. Aliquots were plated on six-well trays of Vero cells for 1 h at 37°C. The inoculum was then replaced with 40% (v v⁻¹) carboxymethylcellulose in DMEM media. HSV-1 plaque assays were incubated for 3–4 days. The monolayers were stained for 1 h with crystal violet stain (Sigma-Aldrich, St. Louis, MO, USA). After removal of the stain, the trays were rinsed with water and dried, and plaques were counted.

All animals were handled in strict accordance with good animal practice as defined by the European directive 63/2010/EU and in accordance with recommendations of the Weatherall report. Four to six-year-old cynomolgus macaques (Macaca fascicularis) were imported from the international accredited breeding facilities from Mauritius (negative for SIV, STLV, herpes B virus, filoviruses, SRV-1, SRV-2, measles, dengue virus and CHIKV) and were housed in a BSL3 facility (Permit Number A 92-032-02), in accordance with Office for Laboratory Animal Welfare (OLAW, USA; #A5826-01) standards at the CEA in accordance with the French national regulation under the number B-92-032-02 for animal use, under the number 2005-69 for macaque breeding. Animals are fed daily and monitored closely by caretakers reporting directly to the veterinarians in charge of the animal facilities. All studies were reviewed and approved by the regional animal care and use committee in accordance with European directive 63/2010/EU: “Comité regional d’éthique pour l’expérimentation animale Ile-de-France Sud”, Fontenay aux Roses, decision #07_012.
Cell lines originally purchased from American Tissue Culture Collection (ATCC), such as human embryonic kidney (HEK 293T, ATCC CRL-3216) and baby hamster kidney (BHK21, ATCC CCL-10) cells, were adhered to recommended ethics approvals and standards.