Penicillin streptomycin

We plated 1000 cells per well in a 96-well plate and incubated them for 24 h with in-media high-glucose DMEM (Microgem, Italy) supplemented with 10% FBS (Microgem, Italy), and 1% penicillin/streptomycin (Microgem, Italy). Tested drugs were added to the media at different concentrations respectively. At the end of the incubation period, the toxicity was evaluated with Cytotoxicity Detection Kit (LDH) (Sigma-Aldrich, Saint Louis, MO, USA) following the manufacturer’s instructions. The untreated sample was the control (100% alive cells) and the percentage of live cells in drug-treated samples was calculated accordingly.

MM cell lines, OPM2 (ACC-50) LP1 (ACC-41), KMS12 (ACC-551) and JJN3 (ACC-541) were purchased from the DSMZ collection of microorganisms; U266 (ATCC^® TIB-196) and H929 (ATCC^® CRL-906) were purchased from the American Type Culture Collection. Cells were cultured in RPMI1640 medium (Euroclone, Pero, Italy) supplemented with 10% fetal bovine serum (Euroclone, Italy), 100 U/mL penicillin/streptomycin (Microgem, Napoli, Italy) and 2 mM l-glutamine (Microgem, Italy).

Human microvascular endothelial cells 1 (HMEC-1) (ATCC® CRL3243™) were cultured in EndoGRO™-MV Complete Culture Media Kit (Millipore) and 1% penicillin/streptomycin (Pan Biotech) and passaged 80-90% confluence.
Dulbecco's Modified Eagle's Medium (DMEM) with phenol red (Sigma-Aldrich) 2% FBS (Microgem), 1% penicillin/streptomycin, and 1% L-glutamine (Microgem) was used for preconditioning and reoxygenation processes.