Macrophages were isolated from C57BL/6 mice (age 8–10 weeks) obtained from the Department of Experimental Biological Models of E.D. Goldberg Institute of Pharmacology and Regenerative Medicine. We performed this research according to EU Directive 2010/63/EU concerning the protection of animals used for scientific purposes, and it was approved by the Animal Care and Use Committee of the Goldberg Research Institute of Pharmacology and Regenerative Medicine, Tomsk NRMC (Protocol No. 171052020 from 05.18.20).
Macrophages were isolated from peritoneal exudate using an EasySep™Biotin Positive Selection Kit and Anti-Mouse F4/80 Antibody (both from StemCell Technologies, Vancouver, BC, Canada). The macrophages were cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO, USA) and supplemented with 10% (v/v) heat-inactivated, endotoxin-free fetal bovine serum (FBS) (Hyclone, GB), 20 mM HEPES (Sigma-Aldrich, St. Louis, MO, USA), 50 mM mercaptoethanol (Sigma-Aldrich, St. Louis, MO, USA), 2 mM L-glutamine (Sigma-Aldrich, St. Louis, MO, USA), and 50 μg/mL gentamycin.
THP-1Blue cells obtained from InvivoGen (San Diego, CA, USA) [45 (link)] and human monocyte–macrophage MonoMac-6 cells (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany) [46 (link)] were cultured at 37 °C in a humidified atmosphere containing 5% CO2, as reported previously.
Macrophages were isolated from peritoneal exudate using an EasySep™Biotin Positive Selection Kit and Anti-Mouse F4/80 Antibody (both from StemCell Technologies, Vancouver, BC, Canada). The macrophages were cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO, USA) and supplemented with 10% (v/v) heat-inactivated, endotoxin-free fetal bovine serum (FBS) (Hyclone, GB), 20 mM HEPES (Sigma-Aldrich, St. Louis, MO, USA), 50 mM mercaptoethanol (Sigma-Aldrich, St. Louis, MO, USA), 2 mM L-glutamine (Sigma-Aldrich, St. Louis, MO, USA), and 50 μg/mL gentamycin.
THP-1Blue cells obtained from InvivoGen (San Diego, CA, USA) [45 (link)] and human monocyte–macrophage MonoMac-6 cells (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany) [46 (link)] were cultured at 37 °C in a humidified atmosphere containing 5% CO2, as reported previously.
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