Flow cytometric detection of endothelial progenitor cells (EPC) in acute coronary syndrome

Mesude Falay 1  and Server Aktaş 2
  • 1 Ankara Numune Trainning and Education Hospital – Department of Hematology, Ankara, Turkey
  • 2 Polatlı State Hospital, Ankara, Turkey
Mesude Falay and Server Aktaş

Abstract

Aim:

The aim of the present study is to establish a moAb combination and a study procedure for the accurate determination of circulating EPC using multicolor flow cytometry, as well as to find out whether there is a relation between disease severity and EPC count by comparing the circulating EPC count between the subgroups of patients presented with acute coronary syndrome.

Materials and methods:

Absolute circulating EPC count was identified in 40 healthy subjects and acute coronary syndrome patients using multicolor flow cytometry with a single-tube panel consisting of CD45, CD31, CD34, CD309 and syto 16 monoclonal antibodies.

Results:

Circulating EPC count was 11.33 (7.89–15.25) cells/μL in healthy control group and 4.80 (0.70–10.85) cells/μL in acute coronary syndrome patients.

Conclusion:

In the present study, which investigated whether there is a difference between EPC counts of STEMI and USAP patients that have presented with ACS, EPC count was lower in both disease groups as compared to healthy controls. First the method must be standardized and then harmonization studies are required in order to use this method as a prognostic factor in cardiovascular diseases.

  • 1.

    Ross R, Glomset J, Harker L. Response to injury and atherogenesis. Am J Pathol 1977;86:675–84.

    • PubMed
    • Export Citation
  • 2.

    Gulati R, Jevremovic D, Peterson TE, Witt TA, Kleppe LS, Mueske CS, et al. Autologous culture-modified mononuclear cells confer vascular protection after arterial injury. Circulation 2003;108:1520–6.

    • Crossref
    • PubMed
    • Export Citation
  • 3.

    Wassmann S, Werner N, Czech T, Nickenig G. Improvement of endothelial function by systemic transfusion of vascular progenitor cells. Circ Res 2006;99:e74–83.

    • PubMed
    • Export Citation
  • 4.

    Asahara T, Murohara T, Sullivan A, Silver M, Van Der Zee R, Li T, et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science 1997;275:964–7.

    • Crossref
    • PubMed
    • Export Citation
  • 5.

    Pearson JD. Endothelial progenitor cells–hype or hope? J Thromb Haemostasis 2009;7:255–62.

    • Crossref
    • Export Citation
  • 6.

    Kocher AA, Schuster MD, Szabolcs MJ, Takuma S, Burkhoff D, Wang J, et al. Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med 2001;7:430–6.

    • Crossref
    • PubMed
    • Export Citation
  • 7.

    Bertolini F, Shaked Y, Mancuso P, Kerbel RS. The multifaceted circulating endothelial cell in cancer: towards marker and target identification. Nat Rev Cancer 2006;6:835–45.

    • Crossref
    • PubMed
    • Export Citation
  • 8.

    Fadini GP, Angelo A. Cell-based methods for ex vivo evaluation of human endothelial biology. Cardiovasc Res 2010;87:12–21.

    • Crossref
    • PubMed
    • Export Citation
  • 9.

    Vasa M, Fichtlscherer S, Aicher A, Adler K, Urbich C, Martin H, et al. Dimmeler number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease. Circ Res 2001;89:E1–7.

    • PubMed
    • Export Citation
  • 10.

    Werner N, Kosiol S, Schiegl T, Ahlers P, Walenta K, Link A, et al. Circulating endothelial progenitor cells and cardiovascular outcomes. N Engl J Med 2005;353:999–1007.

    • Crossref
    • PubMed
    • Export Citation
  • 11.

    Avogaro A, Albiero M, Menegazzo L, de Kreutzenberg S, Fadini GP. Endothelial dysfunction in diabetes the role of reparatory mechanisms. Diabetes Care 2011:34(Suppl 2):285–90.

    • Crossref
    • Export Citation
  • 12.

    Lanuti P, Rotta G, Almici C, Avvisati G, Budillon A, Doretto P, et al. Endothelial progenitor cells, defined by the simultaneous surface expression of VEGFR2 and CD133, are not detectable in healthy peripheral and cord blood. Cytometry A 2016;89:259–70.

    • Crossref
    • PubMed
    • Export Citation
  • 13.

    Fadini GP, Losordo D, Dimmeler S. Critical reevaluation of endothelial progenitor cell phenotypes for therapeutic and diagnostic use. Circ Res 2012;110:624–37.

    • Crossref
    • PubMed
    • Export Citation
  • 14.

    Schmidt-Lucke C, Rossig L, Fichtlscherer S, Vasa M, Britten M, Kämper U, et al. Reduced number of circulating endothelial progenitor cells predicts future cardiovascular events: proof of concept for the clinical importance of endogenous vascular repair. Circulation 2005;111:2981–7.

    • Crossref
    • PubMed
    • Export Citation
  • 15.

    Kissa K, Herbomel P. Blood stem cells emerge from aortic endothelium by a novel type of cell transition. Nature 2010;464:112–5.

    • Crossref
    • PubMed
    • Export Citation
  • 16.

    Masouleh BK, Baraniskin A, Schmiegel W, Schroers R. Quantification of circulating endothelial progenitor cells in human peripheral blood: establishing a reliable flow cytometry protocol. J Immunol Methods 2010;357:38–42.

    • Crossref
    • PubMed
    • Export Citation
  • 17.

    Mariucci S, Rovati B, Bencardino K, Manzoni M, Danova M. Flow cytometric detection of circulating endothelial cells and endothelial progenitor cells in healthy subjects. Int J Lab Hematol 2010;32(1p1):e40–8.

    • Crossref
    • PubMed
    • Export Citation
  • 18.

    Körbling M, Reuben JM, Gao H, Lee BN, Harris DM, Cogdell D, et al. Recombinant human granulocyte–colony-stimulating factor–mobilized and apheresis-collected endothelial progenitor cells: a novel blood cell component for therapeutic vasculogenesis. Transfusion 2006;46:1795–802.

    • Crossref
    • PubMed
    • Export Citation
  • 19.

    Fadini GP, Ozaki T, Taguchi A, Meigs J, Dimmeler S, Zeiher AM, et al. Circulating progenitor cell count for cardiovascular risk stratification: a pooled analysis. PLoS One 2010;5:e11488.

    • Crossref
    • PubMed
    • Export Citation
  • 20.

    Strijbos MH, Kraan J, Sleijfer S, Gratama JW. Potential biomarkers in metastatic gastrointestinal stromal tumors. Clin Cancer Res 2007;13:6244.

    • Crossref
    • PubMed
    • Export Citation
  • 21.

    Madonna R, De Caterina R. Cellular and molecular mechanisms of vascular injury in diabetes – part I: pathways of vascular disease in diabetes. Vasc Pharm 2011;54:68–74.

    • Crossref
    • Export Citation
  • 22.

    Mancuso P, Antoniotti P, Quarna J, Calleri A, Rabascio C, Tacchetti C, et al. Validation of a standardized method for enumerating circulating endothelial cells and progenitors: flow cytometry and molecular and ultrastructural analyses. Clin Cancer Res 2009;15:267–73.

    • Crossref
    • PubMed
    • Export Citation
  • 23.

    Fadini GP, Coracina A, Baesso I, Agostini C, Tiengo A, Avogaro A, et al. Peripheral blood CD34+KDR+ endothelial progenitor cells are determinants of subclinical atherosclerosis in a middle-aged general population. Stroke 2006;37:2277–82.

    • Crossref
    • Export Citation
  • 24.

    Fadini GP, Baesso I, Albiero M, Sartore S, Agostini C, Avogaro A. Technical notes on endothelial progenitor cells: ways to escape from the knowledge plateau. Atherosclerosis 2008;197:496–503.

    • Crossref
    • PubMed
    • Export Citation
  • 25.

    Heeschen C, Lehmann R, Honold J, Assmus B, Aicher A, Walter DH, et al. Profoundly reduced neovascularization capacity of bone marrow mononuclear cells derived from patients with chronic ischemic heart disease. Circulation 2004;109:1615–22.

    • Crossref
    • PubMed
    • Export Citation
  • 26.

    Massa M, Rosti V, Ferrario M, Campanelli R, Ramajoli I, Rosso R, et al. Increased circulating hematopoietic and endothelial progenitor cells in the early phase of acute myocardial infarction. Blood 2005;105:199–206.

    • Crossref
    • PubMed
    • Export Citation
  • 27.

    George J, Goldstein E, Abashidze S, Deutsch V, Shmilovich H, Finkelstein A, et al. Circulating endothelial progenitor cells in patients with unstable angina: association with systemic inflammation. Eur Heart J 2004;25:1003–8.

    • Crossref
    • PubMed
    • Export Citation
  • 28.

    Shintani S, Murohara T, Ikeda H. Mobilization of endothelial progenitor cells in patients with acute myocardial infarction. Circulation 2001;103:2776–9.

    • Crossref
    • PubMed
    • Export Citation
  • 29.

    Thum T, Fraccarollo D, Galuppo P. Bone marrow molecular alterations after myocardial infarction: impact on endothelial progenitor cells. Cardiovasc Res 2006;70:50–60.

    • Crossref
    • PubMed
    • Export Citation
  • 30.

    Dimmeler S, Aicher A, Vasa M, Mildner-Rihm C, Adler K, Tiemann M, et al. HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. J Clin Invest 2001;108:391–7.

    • Crossref
    • PubMed
    • Export Citation
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Turkish Journal of Biochemistry (TJB), official journal of Turkish Biochemical Society, is issued electronically every 2 months. The main aim of the journal is to support the research and publishing culture by ensuring that every published manuscript has an added value and thus providing international acceptance of the “readability” of the manuscripts published in the journal.

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