cEC Enrichment and Enumeration Kit, human ( #130-096-170 )

cEC Enrichment and Enumeration Kit, human

Overview

The cEC Enrichment and Enumeration Kit is designed for the enumeration of mature circulating endothelial cells (cECs) from peripheral blood, based on the expression of CD34 and CD146 as well as the lack of CD45.
To ensure reliable enumeration results, the cEC are enriched during the procedure to increase the sensitivity of the flow cytometric analysis.

The kit contains all reagents for:

red blood cell lysis,
cEC enrichment,
blocking of Fc receptor–mediated non-specific labeling of non-target cells,
enumeration of cECs,
exclusion of dead cells,
isotype controls.

Details

Background information
The number of cECs in peripheral blood in healthy individuals is very low. To obtain reliable enumeration results for these rare cells, the sensitivity of flow cytometric analysis needs to be increased. This is achieved by the enrichment of cECs, thus reducing the number of total events that have to be analyzed.

Downstream applications
Over the past decade, several publications suggested that the number of cECs detected in human peripheral whole blood could be a significant marker for vascular damage in conjunction with various diseases, such as

diabetes¹,
heart disease²,
systemic sclerosis³,
or cancer⁴.

cECs are shed from the damaged vasculature and released into the blood circulation. In the course of disease, increased numbers of cECs can be monitored in the peripheral blood.

Columns

MS Columns or MACSQuant Columns

Figure 1

Leukocytes after exclusion of platelets, dead cells, and debris are shown. Cells were stained with CD34-FITC and CD45-PE and analyzed by flow cytometry using the MACSQuant® Analyzer.

Identification of CD34⁺ cells in the original fraction

Leukocytes after exclusion of platelets, dead cells, and debris are shown. Cells were stained with <a href="/en/PG_595_204_CD34.aspx">CD34-FITC</a> and <a href="/en/PG_595_221_CD45.aspx">CD45-PE</a> and analyzed by flow cytometry using the <a href="http://www.macsquant.com">MACSQuant® Analyzer</a>.

Figure 2

Pre-enriched CD34⁺ cells were stained with the cEC Control Cocktail for isotype control of CD146 staining. Leukocytes after exclusion of platelets, dead cells, and debris are shown. (A) Staining with CD34-FITC and CD45-PE. (B) Dot plot, gated on CD34⁺ cells, shows staining with CD45-PE and Mouse IgG1-APC for isotype control. Cells were analyzed by flow cytometry using the MACSQuant® Analyzer.

Isotype control sample

Figure 3

Pre-enriched CD34⁺ cells were stained with the cEC Staining Cocktail for enumeration of CD146⁺ cells. Leukocytes after exclusion of platelets, dead cells, and debris are shown. (A) Staining with CD34-FITC and CD45-PE. (B) Dot plot, gated on CD34⁺ cells, shows staining with CD45-PE and CD146-APC. Cells were analyzed by flow cytometry using the MACSQuant® Analyzer. For details on cEC number calculation, please view datasheet.

cEC staining sample