Rpmi 1640 l glutamine medium

The HMCL MM1S was acquired from American Type Culture Collection, and JJN3, RPMI-8226, U266, and OPM2 were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen. LINF903, an Epstein–Barr virus-transformed B-cell line established from a healthy individual, was obtained from the National DNA Bank of the University of Salamanca, Spain. MM and LINF903 cell lines were cultured in RPMI 1640–l-glutamine medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% of fetal bovine serum (Sigma-Aldrich) and antibiotics (Gibco Life Technologies, Grand Island, NY, USA). All cells were incubated at 37°C in a 5% CO₂ atmosphere. The presence of mycoplasma was routinely checked with a MycoAlert kit (Lonza, Basel, Switzerland) and only mycoplasma-free cells were used in the experiments.

The human myeloma cell lines, NCI-H929 and MM1S were acquired from ATCC (American Type Culture Collection) and JJN3, RPMI-8226, U266, OPM2 and IM9, from DMSZ (Deuthche Sammlung von Mikroorganismen and Zellkulturen). LINF167, LINF692 and LINF903, Epstein-Barr virus (EBV)–transformed B-cell lines established from 3 healthy individuals, were obtained from the National DNA Bank of the University of Salamanca (Spain). MM and LINF cell lines were cultured in RPMI 1640-L-glutamine medium (Sigma-Aldrich, St Louis, MO) supplemented with 10% of fetal bovine serum (FBS) (Sigma-Aldrich) and antibiotics (Gibco Life Technologies, Grand Island, NY). HeLa and HCT116 were obtained from the ATCC and were cultured in DMEM supplemented with L-glutamine (Sigma-Aldrich), 10% FBS and antibiotics. All cells were incubated at 37°C in a 5% CO₂ atmosphere. The presence of mycoplasma was routinely checked with MycoAlert kit (Lonza, Basel, Switzerland) and only mycoplasma free cells were used in the experiments. Bone marrow (BM) samples were obtained from 5 patients with MM with written informed consent in accordance with the Declaration of Helsinki.