Effects of cell type and culture media on Interleukin-6 secretion in response to environmental particles

Abstract

Cultured lung cells provide an alternative to animal exposures for comparing the effects of different types of air pollution particles. Studies of particulate matter in vitro have reported proinflammatory cytokine signaling in response to many types of environmental particles, but there have been few studies comparing identical treatments in multiple cell types or identical cells with alternative cell culture protocols. We compared soil-derived, diesel, coal fly ash, titanium dioxide, and kaolin particles along with soluble vanadium and lipopolysaccharide, applied to airway-derived cells grown in submerged culture. Cell types included A549, BEAS-2B, RAW 264.7, and primary macrophages. The cell culture models (specific combinations of cell types and culture conditions) were reproducibly different in the cytokine signaling responses to the suite of treatments. Further, Interleukin-6 (IL-6) response to the treatments changed when the same cells, BEAS-2B, were grown in KGM versus LHC-9 media or in media containing bovine serum. The effect of changing media composition was reversible over multiple changes of media type. Other variables tested included culture well size and degree of confluence. The observation that sensitivity of a cell type to environmental agonists can be manipulated by modifying culture conditions suggests a novel approach for studying biochemical mechanisms of particle toxicity.

Cytotoxicity screening of 23 engineered nanomaterials using a test matrix of ten cell lines and three different assays

“…In these cells, we only found an oxidative stress response when we exposed the cells to dispersions of 10 μg cm -2 Carbon Black or TiO 2 3 NMs (Figure 8). Similarly, individual reaction patterns upon exposure to different NMs have been observed in a bronchial and an alveolar epithelial cell line (BEAS-2B, A549) [16]. These findings and our data suggest that even when assessing NM toxicity to cells originating from the same organ, the outcome highly depends on the cell type used.…”

“…Primary cells used were mostly alveolar macrophages which were in some cases more in other less sensitive than immortalized cells [16]. However, most of the studies conducted so far were based on one or two cell lines only.…”

Use of human bronchial epithelial cells (BEAS-2B) to study immunological markers resulting from exposure to PM2.5 organic extract from Puerto Rico

“…It has been reported that bronchial epithelial cell responses can vary depending on specific cell culture conditions including growth media. For example, BEAS-2B grown in KGM supplemented media can be more sensitive to induce IL-6 secretion in response to metals such as V, compared to LHC-9 media (Veranth et al , 2008). Therefore, we emphasize that the cell responses described in our work were obtained using KBM media (without any additional growth supplement).…”

Transient Receptor Potential Vanilloid-1 (TRPV1) Is a Mediator of Lung Toxicity for Coal Fly Ash Particulate Material

“…Six chemically and physically different combustion-derived, soil-derived, and manufactured PM, representative of PM that is found in ambient air, were selected as a test set for identifying TRPV1 agonists, based on their environmental relevance, and variable effects on lung cells and/or tissue, as described by our group and others (Veronesi et al, 1999a,b;Agopyan et al, 2003a,b;Veranth et al, 2004Veranth et al, , 2006Veranth et al, , 2008Smith et al, 2006;. CFA1 is a size-fractionated sample of coal fly ash collected from a power plant in Utah.…”