Isolation and Culture of Multiple Myeloma Cells

PBMC, MM and HS-5 cell lines, primary BMSCs from patients were isolated and cultured as previously described [31 (link), 33 (link), 43 (link)]. Malignant CD138⁺ PCs and healthy B cells were isolated with EasySep™ kits (STEMCELL Technologies, USA), after achieving informed consent according to the declaration of Helsinki. The internal Institutional Board approved the use of human material. SaMMi cell line was obtained from a 81 year old woman with a 20% PCs in the bone marrow. CD138⁺ SaMMi PCs were cultured in DMEM 20% FBS with the addition of IL-6 2.5 ng/ml. Co-cultures of HS-5 or BMSCs with INA-6 or patient derived CD138⁺ PCs were stabilized as described in [33 (link)]. The clinical features of the patients analyzed and the immunophenotype/karyotype of SaMMi are detailed in Tables 1 and 2.