Vero (African green monkey kidney fibroblast) cells and BHK-21 (baby hamster kidney fibroblast) cells were cultured as described previously (Goodman et al. 2014 (link)). Briefly, cells were maintained at 37°C in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies, Grand Island, NY) with 8% fetal bovine serum (FBS; Atlas Biologicals, Fort Collins, CO), 1 mM sodium pyruvate (Life Technologies), 27 mM sodium bicarbonate (Life Technologies), 0.1 mM gentamicin (Lonza, Walkersville, MD), and 1 μM amphotericin B (Sigma-Aldrich, St. Louis, MO). Aedes albopictus mosquito C6/36 cells were maintained at 28°C in DMEM (Life Technologies) with 10% FBS (Atlas Biologicals), 0.1 mM nonessential amino acids (Life Technologies), 1 mM sodium pyruvate (Life Technologies), 9 mM sodium bicarbonate (Life Technologies), and 0.1 mM gentamicin (Lonza). ISE6 tick cells, derived from I. scapularis embryonated eggs, were obtained from Ulrike Munderloh (University of Minnesota) and maintained at 32°C, as previously described (Munderloh et al. 1994 (link)).
Protocol detail
Cell Culture Protocols for Vero, BHK-21, and ISE6 Cells
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Aedes
African green monkey
Amino acids
Amphotericin b
Baby
Biologicals
Cells
Eagle
Eggs
Fibroblast cells
Gentamicin
Hamster
Kidney
Mosquito
Pyruvate
Sodium
Sodium bicarbonate
Tick
Corresponding Organization : Centers for Disease Control and Prevention
Other organizations : The University of Texas Medical Branch at Galveston, University of California, Davis
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Variable analysis
independent variables
- Cell line used (Vero, BHK-21, C6/36, ISE6)
- Not explicitly mentioned
- Cell culture temperature (37°C for Vero and BHK-21, 28°C for C6/36, 32°C for ISE6)
- Culture medium composition (DMEM with varying concentrations of FBS, sodium pyruvate, sodium bicarbonate, gentamicin, and amphotericin B)
- No positive or negative controls were explicitly mentioned in the provided information
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