Ebola Virus Isolation Using Huh-7 Cells has Methodological Advantages and Similar Sensitivity to Isolation Using Other Cell Types and Suckling BALB/c Laboratory Mice

Abstract

Following the largest Ebola virus disease outbreak from 2013 to 2016, viral RNA has been detected in survivors from semen and breast milk long after disease recovery. However, as there have been few cases of sexual transmission, it is unclear whether every RNA positive fluid sample contains infectious virus. Virus isolation, typically using cell culture or animal models, can serve as a tool to determine the infectivity of patient samples. However, the sensitivity of these methods has not been assessed for the Ebola virus isolate, Makona. Described here is an efficiency comparison of Ebola virus Makona isolation using Vero E6, Huh-7, monocyte-derived macrophage cells, and suckling laboratory mice. Isolation sensitivity was similar in all methods tested. Laboratory mice and Huh-7 cells were less affected by toxicity from breast milk than Vero E6 and MDM cells. However, the advantages associated with isolation in Huh-7 cells over laboratory mice, including cost effectiveness, sample volume preservation, and a reduction in animal use, make Huh-7 cells the preferred substrate tested for Ebola virus Makona isolation.

Altered microRNA Transcriptome in Cultured Human Liver Cells upon Infection with Ebola Virus

“…In addition to presenting indications of significant hepatic damage [ 39 ], liver provides the best histopathological features characterizing filovirus infections [ 40 ]. Furthermore, the challenge of isolating emerging viruses appears to be overcome thanks to liver cell lines, like Huh7 cells (hepatocellular carcinoma cells, HCC), which give methodological advantages and comparable sensitivity to viral isolation using other cell types or suckling BALB/c laboratory mice [ 41 ]. The whole-genome expression profiling of liver samples (HepG2 and animal tissue) with EBOV infection has provided precious data on the extensive dysregulation of the metabolism and adaptive immunity through modulation by viral proteins [ 42 , 43 ].…”

Flavonols and dihydroflavonols inhibit the main protease activity of SARS-CoV-2 and the replication of human coronavirus 229E

“…Huh-7 cells, a human hepatocellular carcinoma cell line, are an appropriate to study the replication of different viruses (Nakabayashi et al 1984; Shih et al 1993; Dewi et al 2020; Thongsri et al 2019; Logue et al 2019). This cell line was used for infection and propagation of virus and for testing antiviral activity of compounds.…”

Filoviruses Infect Rhesus Macaque Synoviocytes in Vivo and Primary Human Synoviocytes in Vitro

“…Human hepatocellular carcinoma (Huh-7) cells obtained from Dr. Yoshimi Tsuda (Research Institute for Microbial Diseases, Osaka University, Osaka, Japan) were cultured in Dulbecco's modified Eagle's medium with L-glutamine (Lonza 12-604Q, Williamsport, PA) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, Thermo Fisher Scientific, Waltham, MA) and incubated at 37 C in 5% carbon dioxide as previously described. 38 HFLS and Huh-7 cells were grown for 24 hours in T75 flasks, after which media was removed and replaced with 3 mL of media that contained diluted EBOV/Kik, EBOV/Mak, ma-EBOV/Yam, and MARV/Ang stocks used in nonhuman primate (NHP) studies (multiplicity of infection, 0.01 PFU/cell). Infections were performed in triplicate.…”