Dulbecco s modified eagle s medium dmem

Doxorubicin hydrochloride (DOX; Product Number: D1515; CAS Number: 25316-40-9; formula: C₂₇H₂₉NO₁₁.HCL) was obtained from Sigma–Aldrich Chemicals (St. Louis, MO, USA). DOX has a molecular weight of 579.98 g/mol and is in the form of powder. Methanol (CH₃OH) and chloroform (CHCl₃) were purchased from Sigma–Aldrich. Dulbecco’s Modified Eagle’s medium (DMEM; Lot No: 09221712, GenDEPOT, St. Louis, MI, USA) and 10% (v/v) fetal bovine serum (FBS; Lot No: WB0009) were purchased from GE Healthcare Life Science (Queensland, Australia). HCT116 colon cancer cells were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). TSP-d₄ is a 3-(trimethylsilyl) propionic acid that was purchased from Cambridge Isotope Laboratories in the United States.

The dried roots of E. ebracteolata Hayata were purchased from Kyung Dong market (Seoul, Republic of Korea) and authenticated by Dr. Youngbae Suh, Seoul National University. A voucher specimen (NPRI-16-847) was deposited at the Herbarium of Natural Products Research Institute at Seoul National University. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; #475989), LPS from Escherichia coli (#L2630), Griess reagent (#G4410), G418 (Geneticin; #G8168), and N-tosyl-L-phenylalanyl chloromethyl ketone (TPCK; #T4376) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM; #CM002), penicillin–streptomycin (#CA005), and fetal bovine serum (FBS; #F0600) were purchased from GenDepot (Barker, TX, USA). All antibodies used were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Adipose tissue was collected aseptically from the subgluteal region of a healthy beagle dog under general anesthesia. The tissue was homogenized and incubated with collagenase type 1 enzyme (1 mg/mL, Sigma, St. Louis, MO, U.S.A.) for 2 h at room temperature (16-20°C). The suspension was filtered through a 100 μm nylon mesh then centrifuged at 4°C for 10 min. The supernatant was discarded, and the stromal vascular fraction (SVF) was carefully isolated. The SVF was resuspended with phosphate-buffered saline (PBS) and cultured in a 150 mm Petri dish containing low-glucose Dulbecco's modified Eagle's medium (DMEM; GenDEPOT, Grand Island, NY, U.S.A.) with 10% fetal bovine serum (FBS; Gibco BRL, Grand Island, NY, U.S.A.) and 1% penicillin and streptomycin (PS). Cells were washed with PBS after 24 h of incubation to remove the dead cells and tissue debris, and media were changed. The cells were subcultured when 90% confluence was reached. The third passage (P3) of Ad-MSCs was used in subsequent experiments [21 (link)]. These cells have been previously confirmed for their multiline differentiation capability [16 , 22 (link)].