Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 Feb 1;100(2):565–573. doi: 10.1083/jcb.100.2.565

Novel peripheral blood-derived human cell lines with properties of megakaryocytes

PMCID: PMC2113441  PMID: 2981889

Abstract

For 18 mo, we derived 18 cell lines from 11 donors with various clinical profiles ranging from normal to leukemic. Suspension cultures were initiated with 1 X 10(6) mononuclear blood cells/ml of nutrient medium containing 10% human serum and 10% lectin-stimulated human lymphocyte conditioned medium. The cultures were monitored weekly by morphological analyses of Wright-Giemsa-stained cell preparations. All successful cultures showed a significant decline in viability during the first 3-4 wk with rate "lymphoid" cells observed in mitosis. Within the next 2 wk, the proliferating cells gave rise to a rapidly expanding population of mononuclear cells. As the cultures expanded, cell morphology became heterogeneous with respect to cell size and nuclear ploidy, with an accumulation of giant multinuclear cells that were suggestive of megakarocytes. Even though the cells did not have the classical morphology of mature platelet-forming megakaryocytes, 90% of the cells within a cell line were positive by direct or indirect immunofluorescence for the platelet membrane glycoproteins IIb and IIIa; for surface markers HLA-Dr and B2-microglobulin; for intracellular platelet-derived growth factor and platelet factor IV; and for membrane affinity or binding with serum platelet-derived growth factor and platelet factor IV. These results suggest that a blood precursor cell, most likely a primitive megakaryoblast, was isolated from the peripheral blood and was provided with an optimal culture environment for sustained growth. These cells did not mature to a more differentiated stage, perhaps owing to regulatory factor deficiencies in this in vitro system. The remarkable frequency of obtaining cell lines with megakaryocyte properties from normal peripheral blood and the capacity of some normal donors to repeatedly yield these cell lines make this cell culture system indeed unique by being selective for putative megakaryocyte precursors.

Full Text

The Full Text of this article is available as a PDF (1,018.4 KB).

Selected References

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

  1. Boucher B., Norman C. S. Cold synchronization for the study of peripheral blood and bone marrow chromosomes in leukemias and other hematologic disease states. Hum Genet. 1980;54(2):207–211. doi: 10.1007/BF00278974. [DOI] [PubMed] [Google Scholar]
  2. Brodsky I. Busulphan treatment of polycythaemia vera. Br J Haematol. 1982 Sep;52(1):1–6. doi: 10.1111/j.1365-2141.1982.tb03855.x. [DOI] [PubMed] [Google Scholar]
  3. Brodsky I. Editorial: Role of the megakaryocyte and platelet in the leukemic process in mice and men--a review and hypothesis. J Natl Cancer Inst. 1973 Aug;51(2):329–335. [PubMed] [Google Scholar]
  4. Collins S. J., Gallo R. C., Gallagher R. E. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature. 1977 Nov 24;270(5635):347–349. doi: 10.1038/270347a0. [DOI] [PubMed] [Google Scholar]
  5. Doolittle R. F., Hunkapiller M. W., Hood L. E., Devare S. G., Robbins K. C., Aaronson S. A., Antoniades H. N. Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor. Science. 1983 Jul 15;221(4607):275–277. doi: 10.1126/science.6304883. [DOI] [PubMed] [Google Scholar]
  6. Fauser A. A., Messner H. A. Granuloerythropoietic colonies in human bone marrow, peripheral blood, and cord blood. Blood. 1978 Dec;52(6):1243–1248. [PubMed] [Google Scholar]
  7. Hansen M., Pedersen N. T. Circulating megakaryocytes in blood from the antecubital vein in healthy, adult humans. Scand J Haematol. 1978 Apr;20(4):371–376. doi: 10.1111/j.1600-0609.1978.tb02469.x. [DOI] [PubMed] [Google Scholar]
  8. Jackson C. W., Brown L. K., Somerville B. C., Lyles S. A., Look A. T. Two-color flow cytometric measurement of DNA distributions of rat megakaryocytes in unfixed, unfractionated marrow cell suspensions. Blood. 1984 Apr;63(4):768–778. [PubMed] [Google Scholar]
  9. Kaplan D. R., Chao F. C., Stiles C. D., Antoniades H. N., Scher C. D. Platelet alpha granules contain a growth factor for fibroblasts. Blood. 1979 Jun;53(6):1043–1052. [PubMed] [Google Scholar]
  10. Klein E., Klein G., Nadkarni J. S., Nadkarni J. J., Wigzell H., Clifford P. Surface IgM-kappa specificity on a Burkitt lymphoma cell in vivo and in derived culture lines. Cancer Res. 1968 Jul;28(7):1300–1310. [PubMed] [Google Scholar]
  11. Krishan A. Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol. 1975 Jul;66(1):188–193. doi: 10.1083/jcb.66.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lindahl U., Hök M. Glycosaminoglycans and their binding to biological macromolecules. Annu Rev Biochem. 1978;47:385–417. doi: 10.1146/annurev.bi.47.070178.002125. [DOI] [PubMed] [Google Scholar]
  13. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  14. Martin P., Papayannopoulou T. HEL cells: a new human erythroleukemia cell line with spontaneous and induced globin expression. Science. 1982 Jun 11;216(4551):1233–1235. doi: 10.1126/science.6177045. [DOI] [PubMed] [Google Scholar]
  15. Minowada J., Onuma T., Moore G. E. Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst. 1972 Sep;49(3):891–895. [PubMed] [Google Scholar]
  16. Morgan D. A., Ruscetti F. W., Gallo R. Selective in vitro growth of T lymphocytes from normal human bone marrows. Science. 1976 Sep 10;193(4257):1007–1008. doi: 10.1126/science.181845. [DOI] [PubMed] [Google Scholar]
  17. Reedman B. M., Klein G. Cellular localization of an Epstein-Barr virus (EBV)-associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer. 1973 May;11(3):499–520. doi: 10.1002/ijc.2910110302. [DOI] [PubMed] [Google Scholar]
  18. Scott G. B. Circulating megakaryocytes. Histopathology. 1982 Jul;6(4):467–475. doi: 10.1111/j.1365-2559.1982.tb02743.x. [DOI] [PubMed] [Google Scholar]
  19. Stiles C. D. The molecular biology of platelet-derived growth factor. Cell. 1983 Jul;33(3):653–655. doi: 10.1016/0092-8674(83)90008-9. [DOI] [PubMed] [Google Scholar]
  20. Strausser J. L., Rosenberg S. A. In vitro growth of cytotoxic human lymphocytes. I. Growth of cells sensitized in vitro to alloantigens. J Immunol. 1978 Oct;121(4):1491–1495. [PubMed] [Google Scholar]
  21. Thiagarajan P., Perussia B., De Marco L., Wells K., Trinchieri G. Membrane proteins on human megakaryocytes and platelets identified by monoclonal antibodies. Am J Hematol. 1983 May;14(3):255–269. doi: 10.1002/ajh.2830140307. [DOI] [PubMed] [Google Scholar]
  22. Waterfield M. D., Scrace G. T., Whittle N., Stroobant P., Johnsson A., Wasteson A., Westermark B., Heldin C. H., Huang J. S., Deuel T. F. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature. 1983 Jul 7;304(5921):35–39. doi: 10.1038/304035a0. [DOI] [PubMed] [Google Scholar]
  23. Yunis J. J., Bloomfield C. D., Ensrud K. All patients with acute nonlymphocytic leukemia may have a chromosomal defect. N Engl J Med. 1981 Jul 16;305(3):135–139. doi: 10.1056/NEJM198107163050304. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES