Differential effects of autologous serum on CD34(+) or monocyte-derived dendritic cells

Dendritic cells (DC) with potentially important clinical applications have been generated from human peripheral blood monocytes and CD34(+) cells in t he presence of recombinant cytokines granulocyte-macrophage colony-stimulat ing factor (GM-CSF) + interleukin-4 (IL-4) and GM-CSF + tumor necrosis fact or-alpha (TNF-alpha), respectively. Many of the studies generating DC have included fetal calf serum, which is not desirable due to the risk of immune reactions and infectious disease transmission. Additionally, low DC yields have been reported using serum-free media. In this study, we investigate s upplementing serum-free media with autologous serum and plasma for DC gener ation from monocytes and CD34(+) cells. Our results show that functional DC can be reproducibly obtained in the presence of autologous serum using mon ocytes and CD34(+) cells as the starting populations. However, with the add ition of autologous serum, a differential effect is observed in the phenoty pic characterization of these culture-derived DC. Monocytes cultured for 7 days in X-VIVO 15(TM) serum-free media in the presence of GM-CSF + IL-4 sho wed down-regulation of CD14 with increased expression of HLA-DR, mannose re ceptor, CD80, and CD86, along with highly up-regulated CD1a(+) expression. The addition of autologous serum to serum-free media in monocyte cultures r esulted in a dose-dependent decrease in the CD1a(+) expression generating a distinct subset of CD1a(+/-) cells expressing HLA-DR, mannose receptor, CD 80, and CD86. Upon stimulation with CD40L cells, both monocyte-derived DC s ubsets CD1a(+/-) and CD1a(++) were capable of maturation measured by CD83 a nd CD86 up-regulation. Data suggest the differences in the monocyte-derived DC in serum-free (CD1a(++)) or autologous serum (CD1a(+/-)) supplemented c ultures is of a qualitative nature, rather than quantitative. CD1a(+) and C D14(+) cells expressing HLA-DR, mannose receptor, CD80, and CD86 were gener ated in 7 days from CD34(+) cells in serum-free media. A quantitative effec t was obtained when cultures were supplemented with autologous serum, resul ting in a significant enhancement of CD34-derived DC generated. These resul ts demonstrate generation of DC from two different starting populations usi ng serum-free media that can be enhanced with the addition of autologous se rum. Interestingly, a differential effect was observed in the phenotypic ch aracterization of these culture-derived DC.