Cercopithecus aethiops

The Huh-7 hepatocarcinoma cell line was generously provided by Dr. Sonia Zúñiga (CNB-Spanish National Centre for Biotechnology, Madrid, Spain). This cell line was obtained from a liver tumor in a 57-year-old Japanese male and was used to test HCoV-229E infection [33 (link)]. The Vero cell line (ATCC-CCL81), which was derived from the kidney of an adult African green monkey (Cercopithecus aethiops) was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). All cell lines were cultured in growth medium (GM) containing low-glucose Dulbecco’s modified Eagle medium (Life Technologies, Paisley, UK) supplemented with 5% fetal bovine serum (FBS), 1% (v/v) penicillin G (50 U/mL) and streptomycin (50 µg/mL) (HyClone Laboratories, Logan, UT, USA). Cells were maintained at 37 °C, in a humidified atmosphere of 5% CO₂ (Heraeus HERAcell, Thermofisher. Waltham, MA, USA).

VERO cells, an African Green monkey kidney cell line, were purchased from the European Collection of Authenticated Cell Cultures (ECACC ‐ Code 84121903). VERO cells were cultured in Eagle's minimum essential medium (EMEM) (Lonza, Milano, Italy) supplemented with 2 mM L‐ Glutamine (Lonza, Milano, Italy), 100 units/mL penicillin‐streptomycin mixture (Lonza, Milano, Italy) and fetal bovine serum (FBS) (Euroclone, Pero, Italy) to a final concentration of 5%, at 37°C, in a 5% CO₂ humidified incubator.
VEROE6 cells, an epithelial cell line from the kidney of a normal monkey (Cercopithecus aethiops), were acquired from the American Type Culture Collection (ATCC ‐ CRL 1586).
Huh‐7 cells, an epithelial cell line from Human hepatocellular carcinoma, were kindly provided by the University of Siena (ECACC‐Code 01042712). Both VEROE6 and Huh‐7 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM)‐high glucose (Euroclone, Pero, Italy) supplemented with 2 mM l‐Glutamine (Lonza, Milano, Italy), 100 units/mL penicillin‐streptomycin mixture (Lonza, Milano, Italy) and 10% of FBS, at 37°C, in a 5% CO₂ humidified incubator.
Adherent sub‐confluent cell monolayers of VERO, VERO E6, and Huh‐7 were prepared in growth medium, E‐MEM or D‐MEM high glucose containing 2% FBS in T175 flasks or 96‐well plates for propagation or titration and neutralization tests of SARS‐CoV‐2, respectively.