CD14 MicroBeads can be used for the positive selection or depletion of human monocytes and macrophages from cord blood or PBMCs, as well as pleural, peritoneal, or synovial fluids or from various tissues, such as spleen and lymph node.

Data and images for CD14 MicroBeads, human

Figures

Figure 1

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Separation of CD14
+
cells from PBMCs using CD14 MicroBeads, an MS Column, and a MiniMACS™ Separator.

Figure 1

Separation of CD14
+
cells from PBMCs using CD14 MicroBeads, an MS Column, and a MiniMACS™ Separator.

Figure 2

Monocytes were isolated from PBMCs using CD14 MicroBeads and subsequently cultured for 7 days with IL-4 and GM-CSF to generate immature Mo-DCs and were, finally, cultured for additional 3 days with TNF-α for maturation. Mature Mo-DCs were stained for expression of CD1a, costimulatory molecules (CD80, CD86), CD83 and HLA-DR.
A:
B:
View details

Figure 2

Monocytes were isolated from PBMCs using CD14 MicroBeads and subsequently cultured for 7 days with IL-4 and GM-CSF to generate immature Mo-DCs and were, finally, cultured for additional 3 days with TNF-α for maturation. Mature Mo-DCs were stained for expression of CD1a, costimulatory molecules (CD80, CD86), CD83 and HLA-DR.
View details

Figure 2

Monocytes were isolated from PBMCs using CD14 MicroBeads and subsequently cultured for 7 days with IL-4 and GM-CSF to generate immature Mo-DCs and were, finally, cultured for additional 3 days with TNF-α for maturation. Mature Mo-DCs were stained for expression of CD1a, costimulatory molecules (CD80, CD86), CD83 and HLA-DR.
C:
D:
View details

Figure 2

Monocytes were isolated from PBMCs using CD14 MicroBeads and subsequently cultured for 7 days with IL-4 and GM-CSF to generate immature Mo-DCs and were, finally, cultured for additional 3 days with TNF-α for maturation. Mature Mo-DCs were stained for expression of CD1a, costimulatory molecules (CD80, CD86), CD83 and HLA-DR.
View details

Figure 2

Monocytes were isolated from PBMCs using CD14 MicroBeads and subsequently cultured for 7 days with IL-4 and GM-CSF to generate immature Mo-DCs and were, finally, cultured for additional 3 days with TNF-α for maturation. Mature Mo-DCs were stained for expression of CD1a, costimulatory molecules (CD80, CD86), CD83 and HLA-DR.
E:
View details

Figure 2

Monocytes were isolated from PBMCs using CD14 MicroBeads and subsequently cultured for 7 days with IL-4 and GM-CSF to generate immature Mo-DCs and were, finally, cultured for additional 3 days with TNF-α for maturation. Mature Mo-DCs were stained for expression of CD1a, costimulatory molecules (CD80, CD86), CD83 and HLA-DR.

Specifications for CD14 MicroBeads, human

Overview

CD14 MicroBeads can be used for the positive selection or depletion of human monocytes and macrophages from cord blood or PBMCs, as well as pleural, peritoneal, or synovial fluids or from various tissues, such as spleen and lymph node.

Detailed product information

Background information

The CD14 antigen belongs to the LPS receptor complex. Binding of antibody to CD14 does not trigger signal transduction since CD14 lacks a cytoplasmatic domain. CD14 is strongly expressed on most monocytes and macrophages and weakly on neutrophils and some myeloid dendritic cells.

Downstream applications

The most eminent use of monocytes isolated by MACS® Technology is their
ex vivo
differentiation into monocyte-derived dendritic cells (Mo-DCs). The excellent purity and recovery obtained by using CD14 MicroBeads provide a pure and consistent cell source (fig. 1). Contamination with unwanted cells (e.g. platelets) which may interfere with a controlled differentiation process is markedly reduced, and the isolated monocytes can be induced to differentiate into a homogenous population of immature as well as mature Mo-DCs. Dendritic cells generated from monocytes isolated by MACS® Technology have been used in many different fields of application.
1-6
Apart from the generation of dendritic cells, monocytes are isolated, for example, for
ex vivo
generation of macrophages
7,8
or for studies on cytotoxicity
9
or migration
10
.

Columns

For positive selection: MS, LS, XS, or autoMACS
®
Columns. For depletion: LD, D, or autoMACS Columns.

Reviews for CD14 MicroBeads, human

Great Way to Isolate Monocytes from PBMCs

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CD14 MicroBeads, human (130-050-201)

With this product we achieved successful isolation of monocytes from human PBMCs with >90% purity of CD14+ monocytes and functionally active cells (phagocytosis and cytokine release)

Anti-Human CD14 Microbeads – Quickly Obtain Highly Pure CD14+ Monocytes/Macrophages from PBMC

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CD14 MicroBeads, human (130-050-201)

Our lab requires highly pure immune cell subsets, such as monocytes and macrophages, for our experiments. To purify monocytes and macrophages from human peripheral blood mononuclear cells (PBMC), we regularly employ Miltenyi anti-human CD14 microbeads for magnetic positive selection. This product was initially chosen because of the minimal optimization and straightforwardness of the protocol.

CD14 Microbeads Review

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CD14 MicroBeads, human (130-050-201)

Our laboratory investigates different ways to improve immune responses against HIV, including enhancement of the capacity of effector cells to kill HIV-infected cells. We are currently investigating the capacity of an engineered antibody to trigger ADCC activity in NK cells and monocytes in in vitro assays against an HIV-infected cell line. We use CD14 beads for purification of monocytes and macrophages.

References for CD14 MicroBeads, human

Publications

  1. Pickl, W. F. et al. (1996)
    Molecular and functional characteristics of dendritic cells generated from highly purified CD14
    +
    peripheral blood monocytes.
    J. Immunol. 157: 3850-3859
  2. de Baey, A. and Lanzaveccia, A. (2000) The role of aquaporins in dendritic cell macropinocytosis. J. Exp. Med. 191: 743-747
  3. Salio et al. (2000) Dendritic cell maturation is induced by mycoplasma infection but not by necrotic cells. Eur. J. Immunol. 30: 705-708
  4. Ebner, S. et al. (2002) A novel role for IL-13: Human monocytes cultured in the presence of IL-3 and IL-4 differentiate into dendritic cells that produce less IL-12 and shift the cell responses toward a Tʜ2 cytokine pattern. J. Immunol. 168: 6199-6207
  5. Jefford, M. et al. (2003)
    Functional comparison of DCs generated
    in vivo
    with Flt3 ligand or
    in vitro
    from blood monocytes: differential regulation of function by specific classes of physiologic stimuli.
    Blood 102: 1753-1763
  6. Matsumoto, M. et al. (2003) Subcellular localization of Toll-like receptor 3 in human dendritic cells. J. Immunol. 171: 3154-3162
  7. Hanley, P. J. et al. (2004) Extracellular ATP induces oscillations of intracellular Ca2+ and membrane potential and promotes transcription of IL-6 in macrophages. Proc. Natl. Acad. Sci. U.S.A. 101: 9479-9484
  8. Verreck, F. A. et al. (2004) Human IL-23-producing type 1 macrophages promote but IL-10-producing type 2 macrophages subvert immunity to (myco)bacteria. Proc. Natl. Acad. Sci. U.S.A. 101: 4560-4565
  9. Ryan, E. J. et al. (2002) Dendritic cell-associated lectin-1: a novel dendritic cell-associated, C-type lectin-like molecule enhances T cell secretion of IL-4. J. Immunol. 169: 5638-5648
  10. Vitale, S. et al. (2004) Soluble fractalkine prevents monocyte chemoattractant protein-1-induced monocyte migration via inhibition of stress-activated protein kinase 2/p38 and matrix metalloproteinase activities. J. Immunol. 172: 585-592

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