Toceranib

The chemotherapeutic agents, carboplatin (Sigma-Aldrich, St. Louis, MO, United States), doxorubicin (Sigma-Aldrich), vinorelbine (Sigma-Aldrich), toceranib (Sigma-Aldrich), isophosphoramide mustard (MedChemExpress, Monmouth Junction, NJ, United States), and the active metabolite of ifosfamide were used based on previous publications (2–6 ). Zoledronate (Sigma-Aldrich), carboplatin, and vinorelbine were dissolved in sterile distilled H2O at a final concentration of 10 mM. doxorubicin and toceranib were dissolved in DMSO at a final concentration of 10 mM. Stock solutions were aliquoted and kept at −20°C for long-term storage. Isophosphoramide mustard was dissolved in DMSO at a final concentration of 100 mM. The stock solution of isophosphoramide was aliquoted and kept at −80°C. The stock solutions of carboplatin, doxorubicin, vinorelbine, and toceranib were utilized within 1 year after dissolution. Zoledronate and isophosphoramide mustard were used within 6 months following dissolution. Information on the stability of frozen solutions was only accessible for isophosphoramide mustard, and it remains stable within 6 months.

Ibrutinib was obtained from Selleck Chemicals (Houston, Texas), toceranib from Sigma‐Aldrich (St. Louis, Missouri), masitinib and midostaurin from LC laboratories (Woburn, Massachusetts). Stock solutions for all drugs were prepared by dissolving in dimethyl sulfoxide (DMSO) purchased from Sigma‐Aldrich. RPMI 1640 medium, Iscove's modified Dulbecco's medium (IMDM) and antibiotics (penicillin, streptomycin) were purchased from Lonza (Basel, Switzerland), amphotericin B from PAN‐Biotech (Aidenbach, Germany), fetal calf serum (FCS) from Gibco Life Technologies (Carlsbad, California), ³H‐thymidine from PerkinElmer (Waltham, Massachusetts), collagenase type 2 from Worthington (Lakewood, New Jersey) and trypan blue and 4′,6‐diamidino‐2‐phenylindole (DAPI) from Sigma‐Aldrich. DMSO was used as vehicle‐control in all experiments (corresponding to highest drug concentrations) and showed no effects on growth and activation of canine MCs (not shown).

Fetal bovine serum (FBS), and alamarBlue TM Cell Viability Reagent, were purchased from Thermo Fisher Scientific, USA; vincristine sulfate (1 mg/ml) (Pfizer), doxorubicin hydrochloride (Pfizer), temozolomide (Sigma–Aldrich, USA), panobinostat (Sigma–Aldrich, USA), toceranib (Sigma–Aldrich, USA), fluorouracil (50 mg/ml) (Pfizer), cyclophosphamide (500 mg, Sanfer), methotrexate (25 mg/ml, Pfizer), gemcitabine (1 mg, PISA, S.A de C.V.), and cisplatin (50 mg/50 ml, LEMERLY, S.A de C.V.). DMEM/F12 (Thermo Fisher Scientific, USA), and penicillin/streptomycin (Thermo Fisher Scientific, USA).