Detection of endothelial progenitor cells by flow and laser-scanning cytometry
Dominik Lenz, Karstem Lenk, Volker Adams, Andreas Boldt, Rainer Hambrecht, Attila Tárnok
Abstract
Dominik Lenz, Karstem Lenk, Volker Adams, Andreas Boldt, Rainer Hambrecht, Attila Tárnok
Abstract
Increasing evidence suggests, that endothelial progenitor cells (EPC) play an important role in postnatal neovascularization. The formation of new blood vessels is important in many procedures, e.g. embryogenesis, wound healing, tumor growth and neovascularization of ischemic tissue. Aim of this study was to evaluate an assay which is able to detect EPCs qualitatively as well as quantitatively. This was done by Flo cytometry (FCM) and Laser Scanning Cytometry (LSC). Peripheral blood was drawn out of healthy control persons. In Flow Cytometry mononuclear cells of the peripheral blood, KDR and CD34 double positive cells were defined as precursors of EPCs. Cells from the same specimen were cultured and measured by LSC. While measuring with the LSC it was possible to exclude artifacts o debris by controlling the triggering. The specimen measured with FCM and LSC were examined serologically too, regarding cytokines which usually appear with EPCs (e.g. vascular endothelial growth factor [VEGF]). However, measured results had a good correlation, i.e. a higher amount of precursor cells were accompanied with higher EPC counts.
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Increasing evidence suggests, that endothelial progenitor cells (EPC) play an important role in postnatal neovascularization. The formation of new blood vessels is important in many procedures, e.g. embryogenesis, wound healing, tumor growth and neovascularization of ischemic tissue. Aim of this study was to evaluate an assay which is able to detect EPCs qualitatively as well as quantitatively. This was done by Flo cytometry (FCM) and Laser Scanning Cytometry (LSC). Peripheral blood was drawn out of healthy control persons. In Flow Cytometry mononuclear cells of the peripheral blood, KDR and CD34 double positive cells were defined as precursors of EPCs. Cells from the same specimen were cultured and measured by LSC. While measuring with the LSC it was possible to exclude artifacts o debris by controlling the triggering. The specimen measured with FCM and LSC were examined serologically too, regarding cytokines which usually appear with EPCs (e.g. vascular endothelial growth factor [VEGF]). However, measured results had a good correlation, i.e. a higher amount of precursor cells were accompanied with higher EPC counts.
Key concepts: Flow cytometry, Neovascularization, Progenitor cell, CD34, Peripheral blood mononuclear cell, Vascular endothelial growth factor, Angiogenesis, Cytometry