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. 1988 Dec;9:36–40.

A novel in vitro assay for murine haematopoietic stem cells.

L Eckmann 1, M Freshney 1, E G Wright 1, A Sproul 1, N Wilkie 1, I B Pragnell 1
PMCID: PMC2149103  PMID: 3254725

Abstract

Study of the biology of haematopoietic stem cells is crucially dependent on the availability of suitable in vitro assays. Existing assays have suffered from the fact that they detect small subcompartments of the total stem cell compartment. This limits experiments where it is required to assay a high proportion of stem cells, e.g. the enumeration of stem cell numbers under varying conditions or the identification and purification of stem cell regulators. We describe an in vitro assay which shows macroscopic colony formation and limited self-renewal capacity in vitro. The detected cell (CFU-A) has a low cycling status in normal bone marrow (NBM) and responds to known stem cell regulators. The incidence (100-200 per 10(5) in NBM), the proliferative characteristics under stress and some of the physical properties are similar to stem cells detected by colony formation after transplantation into lethally irradiated recipients (CFU-S). These data indicate that our assay detects a high proportion of haematopoietic stem cells in vitro. This will facilitate experiments on stem cell behaviour which have previously been difficult to conduct.

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Selected References

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

  1. Bradley T. R., Hodgson G. S. Detection of primitive macrophage progenitor cells in mouse bone marrow. Blood. 1979 Dec;54(6):1446–1450. [PubMed] [Google Scholar]
  2. Clark S. C., Kamen R. The human hematopoietic colony-stimulating factors. Science. 1987 Jun 5;236(4806):1229–1237. doi: 10.1126/science.3296190. [DOI] [PubMed] [Google Scholar]
  3. Fialkow P. J., Jacobson R. J., Papayannopoulou T. Chronic myelocytic leukemia: clonal origin in a stem cell common to the granulocyte, erythrocyte, platelet and monocyte/macrophage. Am J Med. 1977 Jul;63(1):125–130. doi: 10.1016/0002-9343(77)90124-3. [DOI] [PubMed] [Google Scholar]
  4. Fialkow P. J., Singer J. W., Raskind W. H., Adamson J. W., Jacobson R. J., Bernstein I. D., Dow L. W., Najfeld V., Veith R. Clonal development, stem-cell differentiation, and clinical remissions in acute nonlymphocytic leukemia. N Engl J Med. 1987 Aug 20;317(8):468–473. doi: 10.1056/NEJM198708203170802. [DOI] [PubMed] [Google Scholar]
  5. Franz T., Löhler J., Fusco A., Pragnell I., Nobis P., Padua R., Ostertag W. Transformation of mononuclear phagocytes in vivo and malignant histiocytosis caused by a novel murine spleen focus-forming virus. Nature. 1985 May 9;315(6015):149–151. doi: 10.1038/315149a0. [DOI] [PubMed] [Google Scholar]
  6. Hollander Z., Shah V. O., Civin C. I., Loken M. R. Assessment of proliferation during maturation of the B lymphoid lineage in normal human bone marrow. Blood. 1988 Feb;71(2):528–531. [PubMed] [Google Scholar]
  7. Johnson G. R., Metcalf D. Pure and mixed erythroid colony formation in vitro stimulated by spleen conditioned medium with no detectable erythropoietin. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3879–3882. doi: 10.1073/pnas.74.9.3879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lahiri S. K., Keizer H. J., van Putten L. M. The efficiency of the assay for haemopoietic colony forming cells. Cell Tissue Kinet. 1970 Oct;3(4):355–362. doi: 10.1111/j.1365-2184.1970.tb00343.x. [DOI] [PubMed] [Google Scholar]
  9. Lala P. K., Johnson G. R. Monoclonal origin of B lymphocyte colony-forming cells in spleen colonies formed by multipotential hemopoietic stem cells. J Exp Med. 1978 Dec 1;148(6):1468–1477. doi: 10.1084/jem.148.6.1468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lord B. I., Mori K. J., Wright E. G. A stimulator of stem cell proliferation in regenerating bone marrow. Biomedicine. 1977 Jul;27(6):223–226. [PubMed] [Google Scholar]
  11. Lord B. I., Mori K. J., Wright E. G., Lajtha L. G. Inhibitor of stem cell proliferation in normal bone marrow. Br J Haematol. 1976 Nov;34(3):441–445. doi: 10.1111/j.1365-2141.1976.tb03590.x. [DOI] [PubMed] [Google Scholar]
  12. Lord B. I., Spooncer E. Isolation of haemopoietic spleen colony forming cells. Lymphokine Res. 1986 Winter;5(1):59–72. [PubMed] [Google Scholar]
  13. Metcalf D., Johnson G. R., Mandel T. E. Colony formation in agar by multipotential hemopoietic cells. J Cell Physiol. 1979 Feb;98(2):401–420. doi: 10.1002/jcp.1040980216. [DOI] [PubMed] [Google Scholar]
  14. Metcalf D. The molecular biology and functions of the granulocyte-macrophage colony-stimulating factors. Blood. 1986 Feb;67(2):257–267. [PubMed] [Google Scholar]
  15. Nakahata T., Ogawa M. Identification in culture of a class of hemopoietic colony-forming units with extensive capability to self-renew and generate multipotential hemopoietic colonies. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3843–3847. doi: 10.1073/pnas.79.12.3843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pragnell I. B., Wright E. G., Lorimore S. A., Adam J., Rosendaal M., DeLamarter J. F., Freshney M., Eckmann L., Sproul A., Wilkie N. The effect of stem cell proliferation regulators demonstrated with an in vitro assay. Blood. 1988 Jul;72(1):196–201. [PubMed] [Google Scholar]
  17. Schofield R. A comparative study of the repopulating potential of grafts from various haemopoietic sources: CFU repopulation. Cell Tissue Kinet. 1970 Apr;3(2):119–130. doi: 10.1111/j.1365-2184.1970.tb00259.x. [DOI] [PubMed] [Google Scholar]
  18. TILL J. E., McCULLOCH E. A. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat Res. 1961 Feb;14:213–222. [PubMed] [Google Scholar]
  19. Tachibana N., Raimondi S. C., Lauer S. J., Sartain P., Dow L. W. Evidence for a multipotential stem cell disease in some childhood Philadelphia chromosome-positive acute lymphoblastic leukemia. Blood. 1987 Nov;70(5):1458–1461. [PubMed] [Google Scholar]
  20. Visser J. W., Bauman J. G., Mulder A. H., Eliason J. F., de Leeuw A. M. Isolation of murine pluripotent hemopoietic stem cells. J Exp Med. 1984 Jun 1;159(6):1576–1590. doi: 10.1084/jem.159.6.1576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wright E. G., Sheridan P., Moore M. A. An inhibitor of murine stem cell proliferation produced by normal human bone marrow. Leuk Res. 1980;4(3):309–314. doi: 10.1016/0145-2126(80)90038-7. [DOI] [PubMed] [Google Scholar]
  22. van den Engh G., Visser J. Light scattering properties of pluripotent and committed haemopoietic stem cells. Acta Haematol. 1979;62(5-6):289–298. doi: 10.1159/000207592. [DOI] [PubMed] [Google Scholar]

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