YILIMART - EPC Enrichment And Enumeration Kit, h

The EPC Enrichment and Enumeration Kit is designed for the enumeration of circulating EPCs (cEPCs) from peripheral blood, cord blood, or leukapheresis products as well as EPCs from bone marrow. To ensure reliable enumeration results, the EPCs are enriched during the procedure to increase the sensitivity of the flow cytometric analysis.

The kit contains all reagents for:

red blood cell lysis,
EPC enrichment,
blocking of Fc receptor–mediated non-specific labeling of non-target cells,
enumeration of EPCs based on expression of CD34, CD133, and CD309,
exclusion of CD14⁺ cells,
exclusion of dead cells,
isotype controls.

Background information

Endothelial progenitor cells (EPCs) are defined by the expression of CD34, CD133, and CD309 (VEGFR-2/KDR)

^1,2

and have been enumerated based on the expression of these markers by several studies. They are considered a measurable parameter for the assessment of risk factors for various diseases

^3–5

as well as for the capacity for vascular repairing. cEPC numbers were correlated with, for example,

certain phases during congestive heart failure⁶ ,
acute myocardial infarction⁷ ,
risk factors for atherosclerosis⁵ ,
risk factors for cardiovascular disease⁸ ,
physical training⁹ ,
cessation of smoking¹⁰ .

The EPC Enrichment and Eunmeration Kit contains everything needed for EPC enrichment followed by enumeration with CD34, CD133/2, CD309, CD14 fluorochrome-conjugated monoclonal antibodies.

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MS or MACSQuant

^®

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Figure 1
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Figure 1

Identification of CD34
⁺
cells and CD133 specificity. Leukocytes after discrimination of debris and platelets and exclusion of dead cells and monocytes are shown. Cells were stained with CD34-FITC and Mouse IgG2b-PE representing the isotype control for CD133 staining. Cells were analyzed by flow cytometry.

Identification of CD34
⁺
cells and CD133 specificity. Leukocytes after discrimination of debris and platelets and exclusion of dead cells and monocytes are shown. Cells were stained with CD34-FITC and Mouse IgG2b-PE representing the isotype control for CD133 staining. Cells were analyzed by flow cytometry.
Figure 2
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Figure 2

Identification of CD34
⁺
CD133
⁺
cells in the pre-enriched EPC sample. Leukocytes after discrimination of cell debris and platelets and exclusion of dead cells and monocytes are shown. Cells were stained with CD34-FITC and CD133/2 (293C3)-PE and analyzed by flow cytometry. The population in R4 was used for further analysis of CD133 and CD309 expression shown in figures 3 and 4.

Identification of CD34
⁺
CD133
⁺
cells in the pre-enriched EPC sample. Leukocytes after discrimination of cell debris and platelets and exclusion of dead cells and monocytes are shown. Cells were stained with CD34-FITC and CD133/2 (293C3)-PE and analyzed by flow cytometry. The population in R4 was used for further analysis of CD133 and CD309 expression shown in figures 3 and 4.
Figure 3
View details

Figure 3

Enumeration of CD34
⁺
CD133
⁺
CD309 (VEGFR-2/KDR)
⁺
EPCs in the pre-enriched EPC sample. Leukocytes after discrimination of debris and platelets, exclusion of dead cells and monocytes are shown. Additionally, data were gated on CD34
⁺
cells. Cells were stained with CD309 (VEGFR-2/KDR)-APC and CD133/2 (293C3)-PE and analyzed by flow cytometry.

Enumeration of CD34
⁺
CD133
⁺
CD309 (VEGFR-2/KDR)
⁺
EPCs in the pre-enriched EPC sample. Leukocytes after discrimination of debris and platelets, exclusion of dead cells and monocytes are shown. Additionally, data were gated on CD34
⁺
cells. Cells were stained with CD309 (VEGFR-2/KDR)-APC and CD133/2 (293C3)-PE and analyzed by flow cytometry.
Figure 4
View details

Figure 4

Isotype control of CD309 staining in the pre-enriched EPC sample. Leukocytes after discrimination of debris and platelets, exclusion of dead cells and monocytes are shown. Additionally, data were gated on CD34
⁺
cells. Cells were stained with CD133/2 (293C3)-PE and Mouse IgG1-APC representing the isotype control for CD309 (VEGFR-2/KDR) staining. Cells were analyzed by flow cytometry.

Isotype control of CD309 staining in the pre-enriched EPC sample. Leukocytes after discrimination of debris and platelets, exclusion of dead cells and monocytes are shown. Additionally, data were gated on CD34
⁺
cells. Cells were stained with CD133/2 (293C3)-PE and Mouse IgG1-APC representing the isotype control for CD309 (VEGFR-2/KDR) staining. Cells were analyzed by flow cytometry.

Selected references
1. Peichev, M. et al. (2000)
  
  Expression of VEGFR-2 and AC133 by circulating human CD34
  ⁺
  cells identifies a population of functional endothelial precursors.
  
  Blood 95: 952-958
2. Rafii, S. and Lyden, D. (2003) Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat. Med. 9: 702-712
3. Hill, J. et al. (2003) Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. N. Engl. J. Med. 348: 593-600
4. Urbich, C. and Dimmeler, S. (2005) Risk factors for coronary artery disease, circulating endothelial progenitor cells, and the role of HMG-CoA reductase inhibitors. Kidney Int. 67: 1672-1676
5. Werner, N. and Nickenig, G. (2006) Influence of cardiovascular risk factors on endothelial progenitor cells: limitations for therapy? Arterioscler. Thromb. Vasc. Biol. 26: 257-266
6. Valgimigli, M. et al. (2004)
  
  CD34
  ⁺
  and endothelial progenitor cells in patients with various degrees of congestive heart failure.
  
  Circulation 110: 1209-1212
7. Massa, M. et al. (2005) Increased circulating hematopoietic and endothelial progenitor cells in the early phase of acute myocardial infarction. Blood 105: 199-206
8. Vasa, M. et al. (2001) Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease. Circ. Res. 89: e1-e7
9. Steiner, S. et al. (2005) Endurance training increases the number of endothelial progenitor cells in patients with cardiovascular risk and coronary artery disease. Atherosclerosis 181: 305-310
10. Kondo, T. et al. (2004) Smoking cessation rapidly increases circulating progenitor cells in peripheral blood in chronic smokers. Arterioscler. Thromb. Vasc. Biol. 24: 1442-1447
Brochures and posters

EPC Enrichment and Enumeration Kit
Scientific posters

Standardized enumeration of human endothelial progenitor cells (EPCs) based on a flow cytometric assay

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EPC Enrichment And Enumeration Kit, h

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Background information

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Figure 1

Figure 1

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Figure 3

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Figure 4

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Selected references

Brochures and posters

Scientific posters

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