biostudies-arrayexpressHeather MaundersHomo sapiensMicroArraySuite 5.0https://www.ebi.ac.uk/biostudies/studies/E-TABM-127Cell cultures from three human non-smoker donors were exposed 1:50 dose (1.84ug/cm2) of diluted mainstream cigarette smoke or filtered air for 1h. Cells from four replicate inserts for each dose/time-point were pooled and placed in TRIzol either immediately after exposure or following a 5h or 23h recovery period after the cultures were returned to an incubator at 37ºC, 5% CO2. The ‘no treatment’ control (NTC) remained in the incubator and received neither smoke nor air. In order to perform robust statistical analyses three independent replicate experiments were performed on each of the three donorsbiostudies-arrayexpressGrowth Protocol - Primary human bronchial epithelial cells were purchased from Cambrex (Walkersville, MD, USA). Cells at passage 2 were seeded into cell culture inserts (Transwell-ClearTM, 6.5mm diameter, 0.4?M pore size; Corning) with 100?l of 0.5 x 105 cells in Bronchial Epithelial Growth Medium (BEGM, Cambrex) per insert and 500?l of basal medium. The following day the apical medium was removed such that the cells were at an air-liquid interface. The basal medium was replaced with 300?l of 50% Bronchial Epithelial Basal Medium (BEBM, Cambrex) in Dulbecco’s Modified Eagles Medium (v:v) containing 0.4% (v:v) bovine pituitary extract, 5?g/ml insulin, 75ng/ml hydrocortisone, 10?g/ml transferrin, 6.5ng/ml thyroxine, 0.5?g/ml epinephrine, 0.5ng/ml epidermal growth factor, 15ng/ml retinoic acid and GA-1000. The medium was replaced daily (Mon – Fri) for the following 23 days during which time the cultures differentiated to form a mucociliary epithelium.Nucleic Acid Extraction - Total RNA was isolated from approximately 50 mg of frozen tissue using TRIZol® reagent according to manufacturer's instructions and purified with QIAGEN® RNeasy® kit following the "RNA cleanup" protocol.Labeling - Labeling of 1 µg of total RNA was carried out following Affymetrix® protocols described in "GeneChip Expression Analysis 2004" available at http://www.affymetrix.com/support/technical/manual/ expression_manual.affxMIAME ScoreRaw DataOrganizationAssays and DataMAGE-TAB FilesArray DesignsFeature Extraction - Title: Affymetrix CEL analysis. Description:MetabolomicsUnknownTranscriptomicsGenomicsProteomicsCigarette smoke is a complex mixture of more than 4,000 constituents. Its effects on cell biology are poorly understood, partly because whole smoke exposure in vitro is technically challenging. To investigate the effects of smoke on cell signaling and function, a three-dimensional air-liquid interface model of tracheobronchial epithelium, grown from primary human lung epithelial cells, was exposed to air or whole mainstream cigarette smoke for 1 h in a purpose-designed chamber. Gene expression profiles were then determined at 1, 6, and 24 h postexposure using Affymetrix HGU133-2 Plus microarrays. Cells from three different donors were used in the study, and the experiment was performed in triplicate for each donor. Genes significantly regulated by smoke, compared with the air control, in all experiments were determined. Genes exhibiting differential expression were assigned to functional categories and mapped to signaling pathways. Effects were observed on many cellular processes including xenobiotic metabolism, oxidant/antioxidant balance, and DNA damage and repair. Notably, there was marked downregulation of the transforming growth factor-beta pathway, which has not been previously reported. This study provides important data on the acute effects of whole cigarette smoke on mucociliary epithelium and may be used to gain a greater understanding of smoke toxicity.transcription profiling by arrayHomo sapiensHuman bronchial epithelial cell transcriptome: gene expression changes following acute exposure to whole cigarette smoke in vitroHeather MaundersMaunders H, Patwardhan S, Phillips J, Clack A, Richter A.falseTranscription profiling of human bronchial epithelial cells from cultures exposed to cigarette smoke or filtered airCell cultures from three human non-smoker donors were exposed 1:50 dose (1.84ug/cm2) of diluted mainstream cigarette smoke or filtered air for 1h. Cells from four replicate inserts for each dose/time-point were pooled and placed in TRIzol either immediately after exposure or following a 5h or 23h recovery period after the cultures were returned to an incubator at 37ºC, 5% CO2. The ‘no treatment’ control (NTC) remained in the incubator and received neither smoke nor air. In order to perform robust statistical analyses three independent replicate experiments were performed on each of the three donors2007-01-15T00:00:00Z2022-03-09T02:59:05.721Z2022-03-09T02:59:05.721ZE-TABM-12717220372EFO_000276810.1152/ajplung.00290.2006