| Overview |
| The CD1c (BDCA-1)+ Dendritic Cell Isolation Kit has been developed for the isolation of CD1c (BDCA-1)+ myeloid dendritic cells (MDCs; type-1 MDCs or MDC1s) from PBMCs. |
| Details |
Background information
The CD1c (BDCA-1) antigen is specifically expressed on dendritic cells, which are CD11chighCD123low and represent the major subset of MDCs in human blood (about 0.6 % of all PBMCs).1–7 CD1c (BDCA-1)+ MDCs show a monocytoid morphology and express myeloid markers such as CD13 and CD33 as well as Fc receptors such as CD32, CD64, and FceRI. Furthermore, they were determined to be CD4+, Lin (CD3, CD16, CD19, CD20, CD56)–, CD2+, CD45RO+, CD141 (BDCA-3)low, CD303 (BDCA-2)–, and CD304 (BDCA-4/Neuropilin-1)–.1 A minor proportion of CD1c (BDCA-1)+ MDCs expresses CD14 and CD11b. CD1c (BDCA-1) is also found on CD1a+ dendritic cells generated ex vivo from monocytes or hematopoietic precursor cells. In blood, apart from MDCs, a subset of B cells also expresses CD1c (BDCA-1). Therefore, the CD1c (BDCA-1)+ Dendritic Cell Isolation Kit includes CD19 MicroBeads for depletion of B cells prior to the enrichment of CD1c (BDCA-1)+ MDCs. In order to discriminate the CD1c (BDCA-1)+ from CD1c (BDCA-1)– CD141 (BDCA-3)++ MDCs, the former have been designated type-1 MDCs (MDC1s).
The CD1c antigen is a member of the CD1 family of proteins that are structurally related to MHC class I proteins and mediate the presentation of non-peptide antigens towards T cells.14
Detailed separation procedure
First, B cells are depleted with CD19 MicroBeads. Then, MDC1s are enriched by magnetic labeling with biotin-conjugated CD1c antibodies and Anti-Biotin MicroBeads.
Downstream applications
Isolated CD1c (BDCA-1)+ MDCs were used to examine the expression of Toll-like receptors4, 5, 12, chemokine receptors3, 5, 8, or new antigens, e.g., DCAL-16 and EMR27. Further, they were isolated for studies on dendritic cell activation4, migration3, cytokine production4, 5, 10, and T cell polarization4, particularly in comparison with monocyte-derived dendritic cells9. The CD1c (BDCA-1)+ Dendritic Cell Isolation Kit was also used for isolation of MDCs from mice engrafted with human CD34+ hematopoietic progenitor cells11, for gene clustering of dendritic cell subsets in blood and tonsils15, for phenotypical analysis of MDCs in PBMCs and synovial fluid of psoriatic arthritis and rheumatoid arthritis patients16, or as an alternative to CD1a MicroBeads for the isolation of epidermal Langerhans cells13.
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| Figure 1 |
| Isolation of CD1c (BDCA-1)+ myeloid dendritic cells isolated from PBMCs by using the CD1c (BDCA-1)+ Dendritic Cell Isolation Kit, a VarioMACS™ Separator, LD and MS Columns. Cell samples of each separation step were stained for CD19 and indirectly for CD1c (BDCA-1) using Anti-Biotin antibodies. |
| PBMCs before separation |
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| Enriched CD1c (BDCA-1)+ dendritic cells after B cell depletion |
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| Isolated CD1c (BDCA-1)+ myeloid dendritic cells |
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| Figure 2 |
| Working scheme for the isolation of CD1c (BDCA-1)+ myeloid dendritic cells from peripheral blood using the CD1c (BDCA-1)+ Dendritic Cell Isolation Kit. |
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