Open AccessAnalysis of human megakaryocytic cells using dual‐color immunofluorescence labeling
Author(s) -
Law Helen K.W.,
Bol Simon J.L.,
Palatsides Manuela,
Williams Neil T.
Publication year - 2000
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/1097-0320(20001201)41:4<308::aid-cyto9>3.0.co;2-n
Subject(s) - platelet , flow cytometry , cd14 , immunofluorescence , microbiology and biotechnology , haematopoiesis , monocyte , platelet membrane glycoprotein , biology , glycoprotein , cell , chemistry , immunology , antibody , stem cell , biochemistry
Background Megakaryocytes are classically identified by their cellular morphology and expression of platelet glycoproteins. Methods In this study, the expression of GPIIIa (CD61) on hemopoietic cells was analyzed by dual‐fluorescence flow cytometry. Results All monocytic cells (CD14+) were shown to coexpress CD61. As the expression of platelet protein on these monocytic cells cannot be reduced by treating the cells with anticoagulant (ethlyenediaminetetraacetic acid [EDTA]), this observation is not simply due to platelet adhesion. When sorted CD61 lo CD14+ cells were studied by light and electron microscopy, platelets or platelet fragments could not be detected on the cell surface. These cells were found to have typical monocytic morphology but no megakaryocytic characteristics. Conclusions This finding demonstrates that without careful definition, the quantitation of megakaryocytic cells will be inappropriately high. A clear and unambiguous criteria for the identification of megakaryocytic cells is described based on the high expression of platelet glycoprotein (e.g., CD61 hi or CD41 hi ) but not the monocyte marker (CD14 neg ). Cytometry 41:308–315, 2000. © 2000 Wiley‐Liss, Inc.