Comparison of Different Culture Microenvironments for the Generation of Dendritic Cells from Peripheral Blood Monocytes of Multiple Myeloma Patients - Preliminary Report

Background. Many protocols regarding the ex vivo generation of dendritic cells (DCs) in culture medium with different protein supplementation have been described. Objectives. This study attempted to generate autologous DCs from peripheral blood mononuclear cells (PBMCs) of multiple myeloma (MM) patients and compare different culture conditions of DC generation. Material and Methods. PBMCs from MM patients were cultured in medium supplemented with 2% human albumin, 10% autologous serum, or 10% allogenic serum from healthy donors in the presence of IL-4 and GM-CSF to generate immature dendritic cells. Maturation of the generated cells was induced by addition of TNF-alpha. Afterwards, the course of apoptosis, phagocytic ability, and the immunophenotype of the generated autologous DCs were analyzed. The concentrations of tumor-derived factors such as IL-6, TGF-beta, VEGF, and IL-10 in serum from the MM patients were also determined. Results. There was a significantly higher percentage of CD1a(+)/CD14(-) cells in the culture with human albumin than in the cultures with addition of autologous or allogenic serum. Moreover, the percentage of Annexin V-positive (AV+) cells in the populations of CD1a(+)/CD14(+) cells and CD1a(+)/CD14(-) cells was significantly higher in human albumin-supplemented culture. Significantly lower percentages of AV(+)/propidium iodide(-) (AV(+)/PI-), AV(+)/PI+, and AV(-)/PI+ cells were found in the culture supplemented with autologous serum than with human albumin. The phagocytosis of FITC-dextran particles by DCs was higher in the cultures supplemented with autologous or allogenic serum than with human albumin. The percentages of CD83(+)/CD1a(+)/HLA-DR+ and CD83(+)/CD1a(-)/HLA-DR+ cells were lower in human albumin-supplemented than in allogenic serum-supplemented culture. Conclusions. These data suggest the possibility of generating autologous DCs from PBMCs of MM patients in medium supplemented with different kinds of serum supplementation. Of note, DC generation progressed in different ways in the examined microenvironments, but supplementation of the cultures with human albumin resulted in the highest percentage of apoptotic cells (Adv Clin Exp Med 2009, 18, 3, 249-259).