Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1979 Jun 1;149(6):1273–1287. doi: 10.1084/jem.149.6.1273

Human megakaryocytes. I. Characterization of the membrane and cytoplasmic components of isolated marrow megakaryocytes

PMCID: PMC2184888  PMID: 571893

Abstract

Human marrow megakaryocytes have been isolated with high purity and yield by processing marrow cells sequentially through density centrifugation and velocity sedimentation. Analysis of the isolated cells for various platelet-associated components by immunofluorescence demonstrated that fibrinogen, plasma factor VIII antigen (factor VIII:AGN) platelet myosin, platelet glycoproteins I and III are present on the membrane and in the cytoplasm of over 90% of marrow megakaryocytes. Parallel studies of human and mouse megakaryocytes and platelets for IgG receptor (FcR), complement receptor type one (CR1) (C3b receptor), complement receptor type two (CR2) (C3d receptor), and Ia antigen by fluorescence and (or) rosette formation methods were performed. FcR were present on most human megakaryocytes and platelets. The Ia antigen was detected on a proportion (10-15%) of human megakaryocytes but it was undetectable on human platelets. CR1 was found on 20-40% of mouse megakaryocytes and also on a proportion of mouse platelets. These differentiation markers may be of use in monitoring megakaryocyte maturation.

Full Text

The Full Text of this article is available as a PDF (3.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abbas A. K., Unanue E. R. Interrelationships of surface immunoglobulin and Fc receptors on mouse B lymphocytes. J Immunol. 1975 Dec;115(6):1665–1671. [PubMed] [Google Scholar]
  2. Behnke O. An electron microscope study of the megacaryocyte of the rat bone marrow. I. The development of the demarcation membrane system and the platelet surface coat. J Ultrastruct Res. 1968 Sep;24(5):412–433. doi: 10.1016/s0022-5320(68)80046-2. [DOI] [PubMed] [Google Scholar]
  3. Cerottini J. C., Brunner K. T. Localization of mouse isoantigens on the cell surface as revealed by immunofluorescence. Immunology. 1967 Oct;13(4):395–403. [PMC free article] [PubMed] [Google Scholar]
  4. Corash L., Tan H., Gralnick H. R. Heterogeneity of human whole blood platelet subpopulations. I. Relationship between buoyant density, cell volume, and ultrastructure. Blood. 1977 Jan;49(1):71–87. [PubMed] [Google Scholar]
  5. Ebbe S. Biology of megakaryocytes. Prog Hemost Thromb. 1976;3:211–229. [PubMed] [Google Scholar]
  6. Ebbe S., Phalen E., D'Amore P., Howard D. Megakaryocytic responses to thrombocytopenia and thrombocytosis in Sl/Sld mice. Exp Hematol. 1978 Feb;6(2):201–212. [PubMed] [Google Scholar]
  7. Forsgren A., Sjöquist J. "Protein A" from S. aureus. I. Pseudo-immune reaction with human gamma-globulin. J Immunol. 1966 Dec;97(6):822–827. [PubMed] [Google Scholar]
  8. Grey H. M., Rabellino E., Pirofsky B. Immunoglobulins on the surface of lymphocytes. IV. Distribution in hypogammaglobulinemia, cellular immune deficiency, and chronic lymphatic leukemia. J Clin Invest. 1971 Nov;50(11):2368–2375. doi: 10.1172/JCI106735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grossi C. E., Webb S. R., Zicca A., Lydyard P. M., Moretta L., Mingari M. C., Cooper M. D. Morphological and histochemical analyses of two human T-cell subpopulations bearing receptors for IgM or IgG. J Exp Med. 1978 May 1;147(5):1405–1417. doi: 10.1084/jem.147.5.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hallberg T., Gurner B. W., Coombs R. R. Opsonic adherence of sensitized ox red cells to human lymphocytes as measured by rosette formation. Int Arch Allergy Appl Immunol. 1973;44(4):500–513. doi: 10.1159/000230956. [DOI] [PubMed] [Google Scholar]
  11. Hoyer L. W. Von Willebrand's disease. Prog Hemost Thromb. 1976;3:231–287. [PubMed] [Google Scholar]
  12. Jackson C. W. Cholinesterase as a possible marker for early cells of the megakaryocytic series. Blood. 1973 Sep;42(3):413–421. [PubMed] [Google Scholar]
  13. Kincade P. W., Paige C. J., Parkhouse R. M., Lee G. Characterization of murine colony-forming B cells. I. Distribution, resistance to anti-immunoglobulin antibodies, and expression of Ia antigens. J Immunol. 1978 Apr;120(4):1289–1294. [PubMed] [Google Scholar]
  14. Levine R. F., Fedorko M. E. Isolation of intact megakaryocytes from guinea pig femoral marrow. Successful harvest made possible with inhibitions of platelet aggregation; enrichment achieved with a two-step separation technique. J Cell Biol. 1976 Apr;69(1):159–172. doi: 10.1083/jcb.69.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Metcalf D., MacDonald H. R., Odartchenko N., Sordat B. Growth of mouse megakaryocyte colonies in vitro. Proc Natl Acad Sci U S A. 1975 May;72(5):1744–1748. doi: 10.1073/pnas.72.5.1744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moore A., Jaffe E. A., Becker C. G., Nachman R. L. Myosin in cultured human endothelial cells. Br J Haematol. 1977 Jan;35(1):71–79. doi: 10.1111/j.1365-2141.1977.tb00564.x. [DOI] [PubMed] [Google Scholar]
  17. Nachman R. L., Hubbard A., Ferris B. Iodination of the human platelet membrane. Studies of the major surface glycoprotein. J Biol Chem. 1973 Apr 25;248(8):2928–2936. [PubMed] [Google Scholar]
  18. Nachman R. L., Jaffe E. A., Weksler B. B. Immunoinhibition of ristocetin-induced platelet aggregation. J Clin Invest. 1977 Jan;59(1):143–148. doi: 10.1172/JCI108612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nachman R. L., Marcus A. J. Immunological studies of proteins associated with the subcellular fractions of thrombasthenic and afibrinogenaemic platelets. Br J Haematol. 1968 Aug;15(2):181–189. doi: 10.1111/j.1365-2141.1968.tb01528.x. [DOI] [PubMed] [Google Scholar]
  20. Nachman R., Levine R., Jaffe E. A. Synthesis of factor VIII antigen by cultured guinea pig megakaryocytes. J Clin Invest. 1977 Oct;60(4):914–921. doi: 10.1172/JCI108846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nachman R., Levine R., Jaffe E. Synthesis of actin by cultured guinea pig megakaryocytes. Complex formation with fibrin. Biochim Biophys Acta. 1978 Sep 21;543(1):91–105. doi: 10.1016/0304-4165(78)90457-9. [DOI] [PubMed] [Google Scholar]
  22. Nakeff A., Maat B. Separation of megakaryocytes from mouse bone marrow by velocity sedimentation. Blood. 1974 Apr;43(4):591–595. [PubMed] [Google Scholar]
  23. Nurden A. T., Caen J. P. Membrane glycoproteins and human platelet function. Br J Haematol. 1978 Feb;38(2):155–160. doi: 10.1111/j.1365-2141.1978.tb01031.x. [DOI] [PubMed] [Google Scholar]
  24. Penington D. G., Lee N. L., Roxburgh A. E., McGready J. R. Platelet density and size: the interpretation of heterogeneity. Br J Haematol. 1976 Nov;34(3):365–376. doi: 10.1111/j.1365-2141.1976.tb03583.x. [DOI] [PubMed] [Google Scholar]
  25. Penington D. G., Olsen T. E. Megakaryocytes in states of altered platelet production: cell numbers, size and DNA content. Br J Haematol. 1970 Apr;18(4):447–463. doi: 10.1111/j.1365-2141.1970.tb01458.x. [DOI] [PubMed] [Google Scholar]
  26. Pfueller S. L., Weber S., Lüscher E. F. Studies of the mechanism of the human platelet release reaction induced by immunologic stimuli. III. Relationship between the binding of soluble IgG aggregates to the Fc receptor and cell response in the presence and absence of plasma. J Immunol. 1977 Feb;118(2):514–524. [PubMed] [Google Scholar]
  27. Pretlow T. G., Stinson A. J. Separation of megakaryocytes from rat bone marrow cells using velocity sedimentation in an isokinetic gradient ficoll in tissue culture medium. J Cell Physiol. 1976 Jul;88(3):317–322. doi: 10.1002/jcp.1040880307. [DOI] [PubMed] [Google Scholar]
  28. Rabellino E. M., Grey H. M., LaForge S., Pirofsky B., Kashiwagi N., Malley A. Antigen heterogeneity of human B and T lymphocytes. J Clin Invest. 1976 Feb;57(2):390–397. doi: 10.1172/JCI108290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rabellino E. M., Ross G. D., Polley M. J. Membrane receptors of mouse leukocytes. I. Two types of complement receptors for different regions of C3. J Immunol. 1978 Mar;120(3):879–885. [PubMed] [Google Scholar]
  30. Rabellino E. M., Ross G. D., Trang H. T., Williams N., Metcalf D. Membrane receptors of mouse leukocytes. II. Sequential expression of membrane receptors and phagocytic capacity during leukocyte differentiation. J Exp Med. 1978 Feb 1;147(2):434–445. doi: 10.1084/jem.147.2.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rabellino E., Colon S., Grey H. M., Unanue E. R. Immunoglobulins on the surface of lymphocytes. I. Distribution and quantitation. J Exp Med. 1971 Jan 1;133(1):156–167. doi: 10.1084/jem.133.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ridell B., Branehög I. The ultrastructure of the megakaryocytes in idiopathic thrombocytopenic purpura (ITP) in relation to thrombokinetics. Pathol Eur. 1976;11(3):179–187. [PubMed] [Google Scholar]
  33. Ross G. D., Jarowski C. I., Rabellino E. M., Winchester R. J. The sequential appearance of Ia-like antigens and two different complement receptors during the maturation of human neutrophils. J Exp Med. 1978 Mar 1;147(3):730–744. doi: 10.1084/jem.147.3.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ross G. D., Polley M. J. Assay for the two different types of lymphocyte complement receptors. Scand J Immunol. 1976 Jun;Suppl 5:99–111. doi: 10.1111/j.1365-3083.1976.tb03861.x. [DOI] [PubMed] [Google Scholar]
  35. Ross R., Glomset J. A. The pathogenesis of atherosclerosis (first of two parts). N Engl J Med. 1976 Aug 12;295(7):369–377. doi: 10.1056/NEJM197608122950707. [DOI] [PubMed] [Google Scholar]
  36. Shortman K., Williams N., Adams P. The separation of different cell classes from lymphoid organs. V. Simple procedures for the removal of cell debris. Damaged cells and erythroid cells from lymphoid cell suspensions. J Immunol Methods. 1972 May;1(3):273–287. doi: 10.1016/0022-1759(72)90005-1. [DOI] [PubMed] [Google Scholar]
  37. Weiss H. J. Platelet physiology and abnormalities of platelet function (first of two parts). N Engl J Med. 1975 Sep 11;293(11):531–541. doi: 10.1056/NEJM197509112931105. [DOI] [PubMed] [Google Scholar]
  38. Williams N., Jackson H., Rabellino E. M. Proliferation and differentiation of normal granulopoietic cells in continuous bone marrow cultures. J Cell Physiol. 1977 Dec;93(3):435–439. doi: 10.1002/jcp.1040930315. [DOI] [PubMed] [Google Scholar]
  39. Williams N., Jackson H., Sheridan A. P., Murphy M. J., Jr, Elste A., Moore M. A. Regulation of megakaryopoiesis in long-term murine bone marrow cultures. Blood. 1978 Feb;51(2):245–255. [PubMed] [Google Scholar]
  40. Winchester R. J., Meyers P. A., Broxmeyer H. E., Wang C. Y., Moore M. A., Kunkel H. G. Inhibition of human erythropoietic colony formation in culture by treatment with Ia antisera. J Exp Med. 1978 Aug 1;148(2):613–618. doi: 10.1084/jem.148.2.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Winchester R. J., Ross G. D., Jarowski C. I., Wang C. Y., Halper J., Broxmeyer H. E. Expression of Ia-like antigen molecules on human granulocytes during early phases of differentiation. Proc Natl Acad Sci U S A. 1977 Sep;74(9):4012–4016. doi: 10.1073/pnas.74.9.4012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Winchester R. J., Wang C. Y., Halper J., Hoffman T. Studies with B-cell allo- and hetero-antisera: parallel reactivity and special properties. Scand J Immunol. 1976;5(6-7):745–757. doi: 10.1111/j.1365-3083.1976.tb03024.x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES