The ARPE-19 human retinal pigment epithelial cell line (American Type Culture Collection [ATCC], Manassas, VA)17 (link) was cultured in 1:1 Dulbecco's modified Eagle's medium (DMEM):F12 medium (Thermo Fisher Scientific-GIBCO, Grand Island, NY) supplemented with heat-inactivated 10% fetal bovine serum (FBS; GE Healthcare-Hyclone, Logan, UT) at 37°C and 5% CO2 in air. Phenotype of cells was verified by confirming the presence of 69 retinal pigment epithelial cell signature transcripts, which are expressed by the ARPE-19 cell line,18 (link) in the RNA sequencing (RNA-seq) transcriptional profile of EBOV- and mock-infected cells (Gene Expression Omnibus [GEO] Accession Number GSE100839). Ebola virus (Zaire ebolavirus, EBOV variant Mayinga) was amplified in Vero C1008 cells (European Collection of Authenticated Cell Cultures [ECACC], Salisbury, UK), cultured with DMEM supplemented with 10% FBS at 37°C and 5% CO2 in air, and titrated by end-point dilution of culture supernatant in fresh Vero C1008 cell monolayers.
Protocol detail
ARPE-19 Cell Culture and Ebola Virus Propagation
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Cell cultures
Cell line
Cells
Cultured medium
Dilution
Ebola virus
Epithelial cell
European
Gene expression
Human
Phenotype
Retinal pigment
Transcriptional
Vero cell
Zaire ebolavirus
Corresponding Organization : South Australian Health and Medical Research Institute
Other organizations : CSIRO Health and Biosecurity, Commonwealth Scientific and Industrial Research Organisation, University College Dublin, EMBL Australia, Emory University, Mulago Hospital, Infectious Diseases Institute
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Variable analysis
independent variables
- Ebola virus (Zaire ebolavirus, EBOV variant Mayinga) infection
- Presence of 69 retinal pigment epithelial cell signature transcripts in the RNA sequencing (RNA-seq) transcriptional profile of EBOV- and mock-infected cells
- ARPE-19 human retinal pigment epithelial cell line
- Culture medium: 1:1 Dulbecco's modified Eagle's medium (DMEM):F12 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS)
- Culture conditions: 37°C and 5% CO2 in air
- Positive control: Mock-infected ARPE-19 cells
- Negative control: Not mentioned
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