Antibiotic antimycotic solution

Human monocytes were purified from the whole blood of de-identified healthy donors using counter-current centrifugal elutriation by the University of Nebraska Medical Center Elutriation Core Facility. Monocytes were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Cytiva) supplemented with 10% fetal bovine serum (FBS), 1% L-glutamine (200 mM; Corning), 1% HEPES (Cytiva), 1% antibiotic-antimycotic solution (Cytiva, USA), 50 µM beta-mercaptoethanol, and 100 ng/mL human macrophage colony-stimulating factor (BioLegend) for 7 days at 37°C to promote macrophage maturation.

MSCs were isolated from the stromal-vascular fraction of subcutaneous adipose tissues of 36 donors (age 42.17 ± 2.07 years; BMI 29.36 ± 1.01 kg/m², for details, see Tables S1 and S3) using enzymatic digestion [8 (link)]. These cells were CD45-/CD73+/CD105+/CD90+/NG2+/PDGFRβ+ [9 (link)]. All donors gave their informed consent. The study was conducted according to the guidelines of the Declaration of Helsinki and human primary cell collection was approved by the Local Ethic Committees of Burdenko Main Military Clinical Hospital (Moscow, Russia) and the Medical Research and Education Center of Lomonosov Moscow State University (IRB00010587, Moscow, Russia) approved the study protocol (#160, 22 July 2019 and #4, 4 June 2018, respectively). Donors medication presented in Table S1. Primary MSCs were cultured in AdvanceSTEM Mesenchymal Stem Cell Media containing a 10% AdvanceSTEM Supplement (HyClone, Cytiva, Marlborough, MA, USA), 1% antibiotic–antimycotic solution (HyClone, Cytiva, Marlborough, MA, USA) at 37 °C in a 5% CO₂ incubator (Binder, Tuttlingen, Germany, CB210). Cells were passaged at 70–80% confluency using Versen solution (Paneco, Moscow, Russia) and HyQTase solution (HyClone, Cytiva, Marlborough, MA, USA).

Dulbecco’s modified Eagle’s medium (DMEM), 2× RPMI 1640 medium, M199 medium, fetal bovine serum (FBS), sodium pyruvate (100 mM), trypsin-EDTA solution, and phosphate-buffered saline were purchased from Welgen, Inc. (Daegu, Korea). Uridine was purchased from Sigma-Aldrich (St. Louis, MO, USA). Basic fibroblast growth factor (bFGF) and heparin were obtained from PeproTech, Inc. (Rocky Hill, NJ, USA). Antibiotic-antimycotic solution and penicillin-streptomycin were purchased from Cytiva (Marlborough, MA, USA). Matrigel and 8-μm pore Transwell filter chambers were purchased from Corning, Inc. (Tewksbury, MA, USA). Qiazol was purchased from Qiagen (Hilden, Germany). All oligo primers were generated by Cosmo Genetech, Co., Ltd. (Seoul, Korea). The antibodies were purchased from the following suppliers: Cell Signaling Technology Inc. (Danvers, MA, USA) and Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). SB-431542 was purchased from Tocris Bioscience (Ellisville, MO, USA). Doxorubicin hydrochloride and ABCB1-specific inhibitor R(+)-verapamil monohydrochloride hydrate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rhodamine 123 and agarose were purchased from Invitrogen (Eugene, OR, USA).