Centrifuge tubes

All chemicals, cell culture media, and supplements for cell culture and submerged exposure were purchased from Gibco/Thermo Fisher Scientific GmBH (Dreieich, Germany), and all chemicals for ALI exposure were obtained from Sigma-Aldrich Chemie GmBH (Taufkirchen, Germany) or Carl Roth GmbH (Karlsruhe, Germany), except for fetal bovine serum (FBS), which was obtained from Thermo Fisher Scientific GmBH (Dreieich, Germany). Cell culture consumables such as plates, flasks, and centrifuge tubes were purchased from Sarstedt (Nuembrecht, Germany) and Greiner (Kremsmünster, Germany). Trans-well plates and inserts were bought from Corning (Amsterdam, The Netherlands). Min-U-Sil5 quartz particles were purchased from U.S. Silica (Katy, TX, USA). Endotoxin contamination was measured via LAL Chromogenic Endotoxin Quantitation Kit (ThermoFisher Scientific, Karlsruhe, Germany). The LDH assay kits were acquired from Promega (Madison, WI, USA) and Roche (Mannheim, Germany). Alamar Blue was obtained from AbD Serotec (Purchheim, Germany). IL-8 ELISAs were purchased from Invitrogen (Waltham, MA, USA).

Four mL blood samples were obtained from patients before and immediately after the surgery into sodium citrate tubes and then were subjected to DNA damage analysis using the comet assay method previously described [39 (link),40 (link),41 ]. Briefly, blood was carefully laid over 4 mL LSM (Biotech, Austria) and tubes were centrifuged (30 min, 1500 rpm, 20 °C). The formed ring of lymphocytes (Figure 5) was transferred to centrifuge tubes (Sarstedt, Austria) and 10 mL of PBS (Sigma Aldrich, Saint Louis, MO, USA) was added. The concentration of cells in the sample was calculated using a Bürker chamber, and samples were centrifuged again (10 min, 1500 rpm, 8 °C). The sediment was re-suspended with PBS buffer (Sigma Aldrich, Saint Louis, MO, USA) and adjusted to a concentration of 1 million cells/mL.

Chemicals and reagents were purchased from Merck KGaA (Darmstadt, Germany) unless otherwise stated, and were of analytical grade. Culture flasks and cryotubes were purchased from VWR (Oslo, Norway), centrifuge tubes from Sarstedt (Oslo, Norway) and Petaka^® G3 LOT (Celartia, Columbus, OH, USA) hypoxic cell culture bioreactors (hereafter entitled “Petakas”) from Tebu-Bio (Roskilde, Denmark). Horse serum and sodium pyruvate were obtained from Thermo Fisher Scientific (Oslo, Norway) and tetrahydrouridine (THU) from AH diagnostics (Oslo, Norway). Reagents and equipment used for liquid chromatography tandem mass spectrometric methods (LC-MS/MS) are described elsewhere [24 (link),25 (link)].