BEAS-2B cells were employed in in vitro studies of iron uptake, ferritin levels, and release of inflammatory mediators. This is an immortalized line of normal human bronchial epithelium derived by transfection of primary cells with SV40 early-region genes. BEAS-2B cells were grown to 90–100% confluence on uncoated plastic 12-well plates in keratinocyte growth medium (KGM; Clonetics) which is essentially MCDB 153 medium supplemented with 5 ng/mL human epidermal growth factor, 5 mg/mL insulin, 0.5 mg/mL hydrocortisone, 0.15 mM calcium, bovine pituitary extract, 0.1 mM ethanolamine and 0.1 mM phosphoethanolamine. Fresh medium was provided every 48 h.
THP1 cells, a monocyte-like cell line, were also used in in vitro investigation. These were cultured in 75-cm2 tissue culture flasks using RPMI-1640 (Invitrogen, Carlsbad, CA, USA) supplemented with 10% serum (Invitrogen, Carlsbad, CA, USA) and gentamicin solution (20 μg/mL; Sigma, St. Louis, MO, USA). Incubations were in RPMI-1640 supplemented with serum and gentamicin.
THP1 cells, a monocyte-like cell line, were also used in in vitro investigation. These were cultured in 75-cm2 tissue culture flasks using RPMI-1640 (Invitrogen, Carlsbad, CA, USA) supplemented with 10% serum (Invitrogen, Carlsbad, CA, USA) and gentamicin solution (20 μg/mL; Sigma, St. Louis, MO, USA). Incubations were in RPMI-1640 supplemented with serum and gentamicin.
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