Targeting Myeloma Cells with NM

MM1.S human MM 63 (link) and 5TGM1 murine MM 64 (link) cell lines, either naïve or carrying Click Beetle Red (CBR) luciferase and Green Fluorescent Protein (GFP) reporters (MM1.S/CBR/GFP and 5TGM/CBR/GFP, respectively), as well as MM1.S/CBR/GFP resistant to RaST and MM1.S/CBR/GFP without CD49d, were generously provided by Dr. DiPersio (Washington University School of Medicine, WUSM, St. Louis). Cells were routinely cultured in complete medium (CM) consisting of Iscove's Modified Dulbecco's Media (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, ThermoFisher Scientific, Waltham, MA) and 50 µg/mL Gentamycin (Thermo Fisher Scientific, Waltham, MA). The cells were routinely washed in phosphate-buffered saline (PBS, Thermo Fisher Scientific, Waltham, MA), pH 7.4. VLA-4 and αvß3 targeted NM were loaded with TC (Bis(cyclopentadienyl)titanium(IV) dichloride, Sigma-Aldrich, St. Louis, MO) 43 , 46 (link). Protein content was routinely measured with Pierce^TM bicinchoninic acid (BCA) assay (Thermo Fisher Scientific Waltham, MA).