Dmem hxa

The human pancreatic ductal adenocarcinoma cell lines BxPC-3 (EP-CL-0042, Elabscience, Houston, TX, USA) and PANC-1 (CRL-1469, ATCC, Manassas, VA, USA), and primary human normal dermal fibroblasts (hNDF) (C-12302, Promocell, Heidelberg, Germany), were cultured at 10,000 cells/cm² for no more than 15 passages in DMEM (DMEM-HXA, Capricorn, Ebsdorfergrund, Germany) or RPMI (RPMI-XA, Capricorn) in the case of BxPC-3, supplemented with 10% Fetal Bovine Serum (FBS) (S1810; Biowest, Nuaillé, France), L-glutamine (X055, Biowest) and Penicillin/Streptomycin (P/S) (L0022, Biowest). Cultures were maintained at 37 °C and 5% CO₂ in a humidified atmosphere.

HEPG2 and HEK293T cells (ATCC HB-8065 and ATCC CRL-3216) were cultured in DMEM High glucose (Gibco 11960 or Capricorn DMEM-HXA) supplemented with 10% FBS (Capricorn FBS-12A9), 2 mM L-glutamine (Sigma G7513) and 1% penicillin-streptomycin (Sigma P0781). H1581 cells (gift from Dr. R.Thomas) were cultured in RPMI (Gibco 21875 or Capricorn RPMI-XA) supplemented with 10% FBS, 2 mM L-glutamine, 1 mM sodium pyruvate (Sigma S8636) and 1% penicillin-streptomycin. Regular mycoplasma test guaranteed all cell lines were mycoplasma negative.

RVFV strain MP12 used for C6/36 cells, insects, and mice (Richard Elliott and Benjamin Brennan, Institute of Infection, Immunity and Inflammation, Centre for Virus Research, University of Glasgow, Glasgow, UK) was propagated using Vero-E6-cells (Collection of Cell Lines in Veterinary Medicine (CCLV), #CCLV-RIE 929, Friedrich-Loeffler-Institute (FLI), Riems, Germany) on a 96-well plate in Dulbecco’s Modified Eagle’s Medium (DMEM, #DMEM-HXA, Capricorn Scientific GmbH, Ebsdorfergrund, Germany) / 5% fetal bovine serum (FBS; #FBS-HI-12A FBS, Capricorn Scientific GmbH, Ebsdorfergrund, Germany) at a temperature of 33 °C in a humidified atmosphere and a CO₂-content of 5%. Supernatant of infected cells was harvested after 3 days with cells showing a cytopathic effect (CPE), characterized by cell lysis in 80% of cells. Additionally, mice and sheep were infected with the virulent RVFV 35/74 strain provided by the virus stock of the FLI, Riems, Germany. The strain was propagated in a cell culture flask of BHK21 cells using Minimum Essential Medium (MEM) supplemented with penicillin–streptomycin and 2% FBS (FLI Bio-Bank, FLI, Riems, Germany) at a temperature of 37 °C and 5% CO₂. Supernatant was harvested, when cells showed 80% CPE. A TCID₅₀ assay, calculated as described by Spearman and Kaerber^{81 (link)}, was used to determine the virus titer.