Kms 12 bm

A panel of 11 myeloma cell lines was used. Four of the cell lines were in-house: OH-2, IH-1, URVIN and KJON, whereas 7 were from other sources: INA-6, CAG, JJN3 and ANBL-6 were kind gifts from Dr M. Gramatzki (University of Erlangen-Nurnberg, Erlangen, Germany), Dr J. Epstein (University of Arkansas for Medical Sciences, Little Rock, AR, USA), Dr J. Ball (University of Birmingham, UK), and Dr D. Jelinek (Mayo Clinic, Rochester, MN, USA), respectively, KMS-12-BM was obtained from DSMZ (Braunschweig Germany), and RPMI-8226 and U266 were from ATCC (Rockville, MD, USA). URVIN, INA-6 and ANBL-6 cells were grown in 10% heat inactivated fetal calf serum (FCS) in RPMI-1640 (RPMI) supplemented with interleukin (IL)-6 (1 ng/mL) (Biosource, Camarillo, CA, USA). CAG, JJN3, KMS-12-BM, RPMI-8226 and U266 were grown in RPMI with 10, 10, 20, 20 or 15% FCS, respectively. OH-2 and IH-1 were maintained in 10% heat-inactivated human serum (HS) (Department of Immunology and Transfusion Medicine, St. Olav's University Hospital, Trondheim, Norway) whereas KJON was maintained in 5% HS, all in RPMI and IL-6 (2 ng/mL). Cells were cultured at 37°C in a humidified atmosphere containing 5% CO₂. For experiments 2% HS in RPMI was used as medium, with IL-6 (1 ng/mL) added for all IL-6 dependent cell lines.

CCLs originating from MM MOLP-8, KMS-12-BM, RPMI-8226, LP-1, OPM-2 and diffuse large B-cell lymphoma (DLBCL) SU-DHL-5 were purchased from DSMZ [German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany], whereas the MM CCL KMM-1 was purchased from JCRB [Japanese Collection of Research Bioresources Cell Bank, Japan]. DLBCL CCL U2932_M was generously provided by Jose A Martinez-Climent [Molecular Oncology Laboratory, University of Navarra Pamplona, Spain].

RPMI-8226, U266, and NCI-H929 MM cell lines were obtained from American Type Cell Culture Collection (Manassas, VA). LP-1, KMS-12-BM, SKMM-2, and OPM-2 were purchased from the DSMZ (Braunschweig, Germany). MM cell lines were maintained in RPMI-1640 media supplemented with 10% fetal bovine serum in a humidified incubator 37(C with 5% CO₂. To establish human MM cells resistant to BZ, RPMI-8226 and U266 cells were continuously cultured in gradually increasing concentrations of BZ (initially 1 nM and increasing in increments of 1 nM over one year) to 50 nM (RPMI-8226) and 20 nM (U266), respectively. Normal peripheral blood mononuclear cells (PBMCs) were purchased from Stemcell Technologies (Vancouver, BC). Primary human MM cells were obtained from the bone marrow of MM patients at the CTRC/UTHSCSA after obtaining informed consent in accordance with an approved IRB protocol. CD138+ cells were selected using beads from Miltenyi Biotec (Auburn, CA).