Detailed separation procedure
The isolation is performed in a two-step procedure. First, B cells and monocytes are magnetically labeled and depleted using a cocktail of CD19 and CD14 MicroBeads. Subsequently, the pre-enriched dendritic cells in the non-magnetic flow-through fraction are magnetically labeled and enriched using a cocktail of antibodies against the dendritic cell markers CD304 (BDCA-4/Neuropilin-1), CD141 (BDCA-3), and CD1c (BDCA-1). The highly pure enriched cell fraction comprises plasmacytoid dendritic cells, CD1c (BDCA-1)
+
type-1 myeloid dendritic cells (MDC1s), and CD1c (BDCA-1)
-
CD141 (BDCA-3)
bright
type-2 myeloid dendritic cells (MDC2s).
B cells and monocytes are depleted in advance because a subpopulation of B cells expresses CD1c (BDCA-1), and monocytes express CD141 (BDCA-3) at low levels.