title: Culture conditions profoundly impact phenotype in BEAS-2B, a human pulmonary epithelial model.
abstract: BEAS-2B, an immortalized, human lung epithelial cell line, has been used to model pulmonary epithelial function for over 30 years. The BEAS-2B phenotype can be modulated by culture conditions that include the presence or absence of fetal bovine serum (FBS). The popularity of BEAS-2B as a model of arsenic toxicology, and the common use of BEAS-2B cultured both with and without FBS, led us to investigate the impact of FBS on BEAS-2B in the context of arsenic toxicology. Comparison of genome-wide gene expression in BEAS-2B cultured with or without FBS revealed altered expression in several biological pathways, including those related to carcinogenesis and energy metabolism. Real-time measurements of oxygen consumption and glycolysis in BEAS-2B demonstrated that FBS culture conditions were associated with a 1.4-fold increase in total glycolytic capacity, a 1.9-fold increase in basal respiration, a 2.0-fold increase in oxygen consumed for ATP production and a 2.8-fold increase in maximal respiration, compared with BEAS-2B cultured without FBS. Comparisons of the transcriptome changes in BEAS-2B resulting from FBS exposure to the transcriptome changes resulting from exposure to 1 μM sodium arsenite revealed that mRNA levels of 43% of the arsenite-modulated genes were also modulated by FBS. Cytotoxicity studies revealed that BEAS-2B cells exposed to 5% FBS for 8 weeks were almost 5 times more sensitive to arsenite cytotoxicity than non-FBS-exposed BEAS-2B cells. Phenotype changes induced in BEAS-2B by FBS suggest that culture conditions should be carefully considered when using BEAS-2B as an experimental model of arsenic toxicity.
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, Arizona, 85724, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, Arizona, 85724, USA.
authors.forename: Walter T
authors.initials: WT
authors.lastname: Klimecki
mesh_terms: D001151:Arsenic / Q000633:toxicity; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004734:Energy Metabolism / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects; D055106:Genome-Wide Association Study; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects*; D010101:Oxygen Consumption / Q000187:drug effects; D020545:Respiratory Mucosa / Q000166:cytology / Q000187:drug effects*
chemical_list: D001151:Arsenic
keywords: BEAS-2B; FBS; arsenic; energy metabolism; gene expression; glycolysis