FLOW CYTOMETRY IN PRECLINICAL TOXICOLOGY/SAFETY ASSESSMENT - Flow Cytometry in Drug Discovery and Development - page 133

FIGURE 6.3 CEC sorting strategy. (a) Sort gate is applied to CD36 cells recovered from
magnetic separation to confirm the absence of cellular events. (b) CD36 þ magnetically
enriched cells with sort gate applied. (c) Postsort analysis after an enrichment sort. (d) Postsort
analysis after second round of sorting for purity. All samples sorted for subsequent gene
expression analysis were of 90% purity.

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Other antigens investigated for CEC/CEP identification included CD31
(PECAM-1), CD54 (ICAM-1), CD105 (endoglin), CD106 (VCAM-1), CD133
(AC133), CD144 (VE-Cadherin), CD146 (MCAM), rat endothelial cell antigen
(RECA)-1, vascular endothelial growth factor receptor (VEGFR)-1, VEGFR-2,
VEGFR-3, and von Willebrand factor (vWF). It is worth mentioning that CD146
is the most widely used primary identifier in human CEC publications. Contrary to
claims in early publications, CD146 expression is not specific to CEC but is found on
some leukocyte subsets (e.g., activated T-cells) [15] and therefore negative selection
of leukocytes is still necessary. It was hoped that some of these antigens (e.g., CD133)
might be useful to distinguish CEC from CEP and provide information about
activation status (e.g., CD144), but results were equivocal. Within the CD36 þ /Lin
population, very few cells were positive for any of the other antigens examined,