Immunofluorescent Identification of Human Megakaryocyte Colonies Using an Antiplatelet Glycoprotein Antiserum

The development of a satisfactory in vitro assay system for human megakaryocyte colony forming progenitor cells has been delayed by the lack of a suitable marker for cells of human megakaryocyte lineage. For this purpose we raised an antiserum directed against a purified human platelet glycoprotein...

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Bibliographic Details
Published in:Blood 1981-02, Vol.57 (2), p.277-286
Main Authors: Mazur, Eric M., Hoffman, Ronald, Chasis, Joel, Marchesi, Sally, Bruno, Edward
Format: Article
Language:English
Subjects:
Antibody Specificity
Antigens, Surface
Blood Platelets - immunology
Bone Marrow Cells
Clone Cells - immunology
Fluorescent Antibody Technique
Glycoproteins - immunology
Humans
Megakaryocytes - immunology
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Summary:The development of a satisfactory in vitro assay system for human megakaryocyte colony forming progenitor cells has been delayed by the lack of a suitable marker for cells of human megakaryocyte lineage. For this purpose we raised an antiserum directed against a purified human platelet glycoprotein preparation. In conjunction with indirect immunofluorescent staining of human bone marrow, this antiserum labeled only platelets, megakaryocytes, and an infrequent population of small mononuclear cells. These small mononuclear cells, not otherwise identifiable as members of the megakaryocyte series, constituted 22.9% of the total fluorescein positive nucleated bone marrow cells. This antiserum was also used to label colonies cultured from human peripheral blood mononuclear cells using a modified plasma clot technique. A mean of 13 fluorescein-labeled colonies were cloned per 106 mononuclear cells cultured. Granulocyte-macrophage and erythroid burst colonies did not label using this method. No augmentation of colony numbers was found with varying concentrations of erythropoietin, human embryonic kidney cell conditioned media (a source of thrombopoietin), or media conditioned by a human T lymphoblast cell line (a source of both colony stimulating and burst promoting activities). Immunofluorescent labeling for platelet glycoproteins is a convenient phenotypic marker for cells of human megakaryocyte lineage useful in the study of in vitro human megakaryocytopoiesis.
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.V57.2.277.277