Heat inactivated fetal calf serum

This study was performed with the approval of the Mayo Clinic Institutional Review Board, and informed consent was obtained in accordance with the Declaration of Helsinki. The MC-B11/14 HMCL was established from the BM aspirate obtained following the patient’s second relapse. The BM aspirate was processed according to previously described procedures [10 (link)]. At onset, CD138⁺ cells were cultured in Iscove’s modified Dulbecco medium with GlutaMAX (Invitrogen, Carlsbad, CA), supplemented with 50 U/mL penicillin G, 50 μg/mL streptomycin (Invitrogen), 10% heat-inactivated fetal calf serum (Atlanta Biologicals, Norcross, GA), 1 ng/mL interleukin-6 (IL-6; kindly provided by Novartis, Basel, Switzerland) and 10 ng/mL insulin-like growth factor-I (IGF-I; Sigma-Aldrich, St. Louis, MO). The MC-B11/14 HMCL is routinely cultured under these same conditions. This HMCL is responsive to both IL-6 and IGF-I and addition of both cytokines has an additive effect on cell proliferation; thus, the cells are routinely cultured with both cytokines to maintain optimal in vitro cell line passaging. The MC-B11/14 HMCL has been tested for the presence of Epstein Barr virus (EBV) using PCR methodology and was determined to be negative (data not shown). All studies used MC-B11/14 cells that had been in continuous culture for less than three months.