Bronchial epithelial singlequots kit

Beas-2B, A549, H358, H2122, H1975, HCC827, PC9, and H1299 were obtained from the American Type Culture Collection (Manassas, VA, USA) or Korean Cell Line Bank (Seoul, Korea). Beas-2B was maintained in Bronchial Epithelial Cell Growth Basal Medium (BEBM) (Cat. No. CC-3170, Clonetics^®, Lonza Inc., Walkersville, MD, USA) with Bronchial Epithelial SingleQuots kit (Cat. No. CC-4175, Lonza Inc., Walkersville, MD, USA) as supplementary additives at 37 °C containing 5% CO₂. Other cells were maintained in RPMI-1640 medium (Corning, Manassas, VA, USA) supplemented with 10% FBS (Corning) and 1X penicillin–streptomycin (Invitrogen, Carlsbad, CA, USA) at 37 °C containing 5% CO₂.

For the nano-DESI solvent, HPLC-MS grade methanol (BDH Chromanorm Solvents, VWR International AB, Stockholm, Sweden), deionized water (18.2 MΩ, Milli-Q, Millipore, Solna, Sweden), and formic acid (98-100%, Merck, Darmstadt, Germany) were used. Interleukine-13 and histamine were purchased (Sigma-Aldrich, Stockholm, Sweden), and budesonide and terbutaline were extracted from commercially available products. The medium used for growing the cells consisted of 50 mL of Dulbecco’s Modified Eagle Medium (500 mL of D-MDM, 5 mL of 100 mM Minimum Essential Medium Sodium pyruvate (Gibco, MEM NaPyr, Fisher Scientific, Gothenburg, Sweden), 5 mL of 200 mM L-Glutamine, 5 mL of 100× Minimum Essential Medium Non-Essential Amino Acids (Gibco MEM NEA, Fisher Scientific, Gothenburg, Sweden), and 50 mL ALI medium (250 mL of Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza Bioscience, Stockholm, Sweden), 1 vial of Bronchial Epithelial SingleQuots Kit (BEGM, Lonza Bioscience, Stockholm, Sweden), and 500 µL 1.5 mg/mL BSA), with an addition of 50 µL 0.1 mM retinoic acid. All the other reagents were purchased from Thermo Fisher, Waltham, MA, USA.