biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsShinkichi Ishikawatranscription profiling by arrayHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-6471Cigarette smoke (CS) is an aerosol containing more than 6,000 chemicals and one of the risk factor in the development of chronic inflammatory lung disease. To evaluate biological effect of CS on human respiratory tract, organotypic bronchial epithelial cultures can be used to replicate in vivo tissue conditions. The MucilAir organotypic bronchial epithelial cultures were exposed to mainstream aerosols from the 3R4F cigarette and a novel tobacco vapor product (NTV), which we recently developed, using a Vitrocell exposure system. This system consists of three steps: the generation of CS, dilution, and exposure to an air-liquid interface cultured cells in a specially designed module. This exposure scenario mimics CS exposure in the human airway (i.e. direct aerosol exposure to the apical surface of air-liquid interface-cultured cells), We found a dose-dependent increase in the number of differentially expressed genes following 3R4F cigarette smoke exposure, compared with expression in air-exposed controls. In contrast, no changes were detected following exposure to NTV vapor.biostudies-arrayexpressSample Collection - The organotypic human bronchial epithelial cultures MucilAir (product code EP01, Epithelix Sarl, Geneve, Switzerland) were used in this study. The cells were incubated at 37°C with a 5% CO2 atmosphere for acclimation for more than 10 days before use. Then the cells were exposed to mainstream aerosols of 3R4F cigarette or novel tobacco vapor product (NTV) using a VC 10 smoking robot and Vitrocell 12/3 module. In the first exposure experiment, Two, and four 3R4F cigarettes were smoked in accordance with the Health Canada Intensive regime (HCI: 55-mL puffs of 2 sec in duration every 30 sec), and whole smoke was released into the mixing device in 8.0-sec exhausts followed by 1.0 L/min dilution flow. Diluted smoke was introduced into the exposure chamber with 5 mL/min flow controlled using mass flow controllers. Clean air was used as the control. Forty-eight hours after the exposure, mRNA was extracted for transcriptomic analysis. In the second exposure experiment, one 3R4F cigarette and one capsule of NTV were smoked in accordance with HCI regime, and whole aerosol was released into the mixing device in 8.0-sec exhausts followed by 0.0, 1.0 or 3.0 L/min dilution flow. Clean air was used as the control. Forty-eight hours after the exposure, mRNA was extracted for transcriptomic analysis.Hybridization - The biotin-labeled RNA was Hybridized on Affymetrix GeneChip Human Genome U133 Plus 2.0 Array using GeneChip® Hybridization, Wash and Stain Kit.Scaning - Command Console Software (Affymetrix) was used to automatically grid the DAT files and create the CEL files (probe cell intensity data).Nucleic Acid Extraction - Total RNA was isolated and purified using the Qiagen RNeasy® Mini Kit (Qiagen, Hilden, Germany).Labeling - Biotinylated cRNA was generated using GeneChip® 3′IVT PLUS Reagent Kit.MIAME ScoreRaw DataOrganizationAssays and DataMAGE-TAB FilesArray DesignsShinkichi IshikawafalseOrganotypic human bronchial epithelial cultures (MucilAir) exposed to mainstream aerosols from the 3R4F cigarette and a novel tobacco vapor productCigarette smoke (CS) is an aerosol containing more than 6,000 chemicals and one of the risk factor in the development of chronic inflammatory lung disease. To evaluate biological effect of CS on human respiratory tract, organotypic bronchial epithelial cultures can be used to replicate in vivo tissue conditions. The MucilAir organotypic bronchial epithelial cultures were exposed to mainstream aerosols from the 3R4F cigarette and a novel tobacco vapor product (NTV), which we recently developed, using a Vitrocell exposure system. This system consists of three steps: the generation of CS, dilution, and exposure to an air-liquid interface cultured cells in a specially designed module. This exposure scenario mimics CS exposure in the human airway (i.e. direct aerosol exposure to the apical surface of air-liquid interface-cultured cells), We found a dose-dependent increase in the number of differentially expressed genes following 3R4F cigarette smoke exposure, compared with expression in air-exposed controls. In contrast, no changes were detected following exposure to NTV vapor.2018-02-09T00:00:00Z2022-03-07T02:00:16.212Z2022-03-07T02:00:16.212ZE-MTAB-6471EFO_0002768EFO_0003814EFO_0002944EFO_0003813EFO_0005518EFO_0003815