Adrenal Cortical Cell Culture Infection

Primary human adrenal cortical cells (HAdCCs; ScienCell, Carlsbad, CA, USA) were used; cell type was confirmed by an immunofluorescence antibody assay showing the presence of glial fibrillary acidic protein (GFAP; present in adrenal cells in the cortex) and absence of tyrosine hydroxylase (TH; present in adrenal chromaffin cells in the medulla). HAdCCs were seeded at 5000 cells/cm² in mesenchymal medium containing 5% fetal bovine serum (FBS), 1% mesenchymal stem cell growth supplement, and 1% 100X penicillin-streptomycin (ScienCell). After 24 h, media was changed to media with 0.1% FBS and 1% 100X penicillin-streptomycin and was replenished every 72 h for 7 days, establishing quiescence (qHAdCC). On day 7, uninfected (mock) or VZV-infected qHAdCCs (60 plaque-forming units/cm²; VZV Gilden strain [GenBank accession number MH379685] [7 (link)]) were added to qHAdCCs. For analysis, cells were harvested at 1, 2, or 3 days post-infection (DPI), and conditioned supernatants were collected at 3 DPI.

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Niemeyer C.S., Mescher T., Bubak A.N., Medina E.M., Hassell JE J.r, & Nagel M.A. (2022). VZV Infection of Primary Human Adrenal Cortical Cells Produces a Proinflammatory Environment without Cell Death. Viruses, 14(4), 674.

Publication 2022

HAdCCs mesenchymal stem cell growth supplement

Adrenal cortical Antibody Cells Chromaffin cells Cortex Fetal bovine serum Gfap Human Immunofluorescence assay Infection Medulla Mesenchymal Mesenchymal stem cell Penicillin Plaque Strain Streptomycin Supplement Tyrosine hydroxylase

Corresponding Organization : University of Colorado Denver

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Variable analysis

independent variables

Presence or absence of VZV infection (mock or VZV-infected qHAdCCs)

dependent variables

Cell harvesting at 1, 2, or 3 days post-infection (DPI)
Conditioned supernatant collection at 3 DPI

control variables

Cell type (primary human adrenal cortical cells, HAdCCs)
Cell seeding density (5000 cells/cm^2)
Culture media composition (mesenchymal medium with 5% FBS, 1% mesenchymal stem cell growth supplement, and 1% penicillin-streptomycin)
Media change to 0.1% FBS and 1% penicillin-streptomycin after 24 h
Media replenishment every 72 h for 7 days to establish quiescence (qHAdCC)
VZV strain (Gilden strain [GenBank accession number MH379685])
VZV infection dose (60 plaque-forming units/cm^2)

negative control

Uninfected (mock) qHAdCCs

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