Abstract
The bone marrow is the primary site of hematopoiesis in adults. Accurate in vitro models of hematopoietic regulation and function should reflect the various hormonal and environmental regulators that act on developing hematopoietic cells in the marrow. Some of the regulatory elements acting in vivo are maintained when bone marrow cells are cultured in vitro under suitable culture conditions. Modifications of techniques initially described by Dexter (1; see Chapter 16) for culture of murine bone marrow led to the establishment of an in vitro system that supports human hematopoietic cells (2,3). Functional hematopoiesis in these cultures was dependent on the formation, over 3–4 wk, of an adherent layer of cells derived from structural elements of bone marrow. This layer, termed “stromal” layer because of its likely derivation from the supportive elements in the marrow, has been extensively characterised (4). Stromal cells have been shown to exert positive and negative regulatory effects on hematopoietic cells through cell-cell interaction and by the secretion of cytokines (5,6).
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References
Dexter, T. M., Allen, T. D., and Lajtha, L. G. (1977) Conditions controlling the proliferation of haemopoietic stem cells in vitro. J. Cell Physiol. 91, 335–344.
Gartner, S. and Kaplan, H. S. (1980) Long-term culture of human bone marrow cells. Proc. Natl. Acad. Sci. USA 77, 47–56.
Wright, D. G. and Greenberger, J. S., eds. (1984) Long Term Bone Marrow Culture. Liss, New York.
Singer, J. W., Keating, A., and Wight, T. H. (1985) The human hematopoietic microenvironment, in Advances in Haematology, vol. 4 (Hoffbrand, V., ed.) Churchill Livingstone, London, pp. 1–24.
Keating, A. and Gordon, M. Y. (1988) Hierarchial organization of hematopoietic microenvironments. Leukemia 2, 766–769.
Clark, B. R., Gallagher, J. T., and Dexter, T. M. (1992) Cell adhesion in the stromal regulation of haemopoiesis. Balliere’s Clin. Haematol. 5, 619–652.
Testa, N. G. and Molineux, G. (1993) Haemopoiesis: A Practical Approach. IRL Press, Oxford, UK.
Eaves, C. J., Sutherland, H. J., Voomaski, C., Lansdorp, P. M., Szilvassy, S. J., Fraser, C. C., Humpries, R. K., and Barnett, M. J. (1992) The human hematopoietic stem cell in vitro and in vivo. Blood Cells 18, 301–307.
Simmons, P. J., Masinovsky, B., Longencker, B. M., Phillips, G. L., Eaves, A. C., Berenson, R., Torok-Storb, B., and Gallatin, W. M. (1992) Vascular cell adhesion molecule-1 expression by bone marrow stromal cells mediates the binding of hematopoietic progenitor cells. Blood 80, 388–395.
Holdrinet, R. S. G., von Egmond, J., Wessels, J. M., and Haanen, C. (1980) A method of quantification of peripheral blood admixture in bone marrow aspirates. Exp. Hematol. 8, 103–107.
Brandwein, J., Dube, I., Laraya, P., and Keating, A. (1992) Maintenance of Philadelphia chromosome-positive progenitors in long-term marrow cultures from patients with advanced chronic myeloid leukemia. Leukemia 6(6), 556–561.
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© 1997 Humana Press Inc., Totowa, NJ
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Clark, B.R., Jamieson, C., Keating, A. (1997). Human Long-Term Bone Marrow Culture. In: Pollard, J.W., Walker, J.M. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology™, vol 75. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-441-0:249
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DOI: https://doi.org/10.1385/0-89603-441-0:249
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-441-9
Online ISBN: 978-1-59259-561-7
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