Dendritic cells (DCs) form a critical interface between innate and adaptive immunity. DCs continuously sample their environment for antigens by means of endocytosis. Referred to as an antigen-presenting cell (APC), DCs capture, process and present these antigens to naïve T-helper cells to initiate an immune response. There is great heterogeneity in DC phenotypes due to their stage of differentiation or according to their response and interaction with cytokines and/or growth factors in their microenvironment. This can make isolating a specific subset of DCs challenging. Additionally, each subset is rare in number, compounding the difficulty in isolating these cells. Culturing of CD14+ monocytes in the presence of specific growth factors and cytokines can induce differentiation into immature DCs (iDC), known as monocyte-derived immature dendritic cells (MoDCs). Cultured immature dendritic cells remove the phenotype heterogeneity issue as well as the rarity of these cells.

Human peripheral blood monocyte-derived immature dendritic cells are derived from negatively selected monocytes. Non-monocytes are labeled and depleted from the peripheral blood mononuclear cell population using immunomagnetic particles leaving purified, untouched monocytes. Untouched monocytes are cultured for 4-5 days in culture medium containing 10% FBS plus GM-CSF and IL-4. After culture, cells are checked for the expression of CD11c, HLA-DR, and CD83 and the lack of CD14 expression by flow cytometry before cryopreservation.

Cells were obtained using Institutional Review Board (IRB) approved consent forms and protocols.