YILIMART - CD14 MicroBeads, human

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.

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.

Figure 1

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:

C:	D:

E:

Figure 2
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Figure 2

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

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

Selected references
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
Brochures and posters

Human myeloid cells from peripheral blood
Scientific posters

Monocyte-derived DC generation for cancer vaccines
Customer reports and application notes

MACS
^®
Technology facilitates mixed lymphocyte reaction

From monocyte isolation to DC characterization - Generation of Mo-DCs

Effector memory T helper cells secrete IFN-γ upon stimulation with cytokines: a role in chronic inflammation. Sattler et al. (2011) MACS&more 13(1).

Highly purified peripheral blood γ/δ T cells isolated by MACS
^®
Technology respond to NOD2 ligand. Marischen et al. (2013) MACS&more 15(1).
Special protocols

Isolation of CD4
⁺
T helper cells from human PBMCs

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CD14 MicroBeads, human - lyophilized

For research use only

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Background information

Downstream applications

Columns

Figure 1

Figure 2

Figure 2

Selected references

Brochures and posters

Scientific posters

Customer reports and application notes

Special protocols

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