Qiagen RT-PCR array Dimethylamin validation experiment

This study compares the dose dependent transcriptome data obtained for the volatile compound dimethylamine, which has been exposed in an in vitro system via air-liquid-interface (ALI) exposure to a pulmonary cell line (A549 cells). A549 cells were exposed to dimethylamine under ALI-conditions in 12-well plates (P.R.I.T.® ExpoCube®, Ritter et al. 2018 ). Exposure took place for 60 minutes by applying exposure flows of 3 ml/min per 1cm² ALI culture. The experimental design was based on 3 groups including 4 cultures / plate being exposed to the test substance, 4 cultures / plate being exposed to clean air as exposure control and the remaining wells / plate as non-exposure controls (no application of exposure flow). Vapor concentrations were set up by controlled in-line evaporation of the liquids using an impinger and dilution with clean air. Concentrations were monitored online using quantitative FT-IR analysis (Gasmet DX4000). High dose (HD) = 81.7 ppm, mid dose (MD) = 41.1 ppm, low dose (LD) = 14.9 ppm. Exposure for one hour was followed by a 23hr recovery period before the next exposure started. Within a 72hr interval a total of 3 exposure and recovery periods were accomplished without medium change in between. For each exposure condition 3 biological replicates where tested, each was based on 2 pooled wells.