CCHFV Propagation and Culture

Huh7 and SW13 cells were propagated in Dulbecco’s modified Eagle’s medium with Earle’s salts (Corning), both medias were supplemented with 10% fetal bovine serum (FBS; Gibco), 1% penicillin/streptomycin (Gibco), 1% sodium pyruvate (Sigma-Aldrich), 1% l-glutamine (HyClone), and 1% Hepes (Gibco). Minimally passaged CCHFV strain Afg09-2990 (39 (link)) or strain IbAr 10200 (USAMRIID collection) were used for all experiments, as indicated. Strain Afg09-2990 was previously passaged three times in Vero cells and then propagated at USAMRIID two times in Huh7 cells. Strain IbAr 10200 was passaged, as previously described (25 (link)). The viruses were collected from clarified cell culture supernatants and stored at −80°C. All CCHFV work was handled in biosafety level 4 (BSL-4) containment at USAMRIID.

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Golden J.W., Shoemaker C.J., Lindquist M.E., Zeng X., Daye S.P., Williams J.A., Liu J., Coffin K.M., Olschner S., Flusin O., Altamura L.A., Kuehl K.A., Fitzpatrick C.J., Schmaljohn C.S, & Garrison A.R. (2019). GP38-targeting monoclonal antibodies protect adult mice against lethal Crimean-Congo hemorrhagic fever virus infection. Science Advances, 5(7), eaaw9535.

Publication 2019

Earle’s salts penicillin/streptomycin sodium pyruvate Hepes

Cell culture Cells Eagle Glutamine Hepes Penicillin Pyruvate Salts Sodium Strain Streptomycin Vero cells Viruses

Corresponding Organization : United States Army Medical Research Institute of Infectious Diseases

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Variable analysis

independent variables

Huh7 and SW13 cell lines
CCHFV strain Afg09-2990 and strain IbAr 10200

dependent variables

Virus propagation and growth

control variables

Dulbecco's modified Eagle's medium with Earle's salts (Corning)
10% fetal bovine serum (FBS; Gibco)
1% penicillin/streptomycin (Gibco)
1% sodium pyruvate (Sigma-Aldrich)
1% l-glutamine (HyClone)
1% Hepes (Gibco)

controls

Positive control: Vero cells used for initial passaging of CCHFV strain Afg09-2990
Negative control: Not explicitly mentioned

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