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Stromal components in rat bone marrow frozen sections compared to long-term rat bone marrow cultures

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Abstract

The haematopoietic microenvironment is believed to play an important role in controlling the haematopoietic process. Ultrastructural studies have shown that the haematopoietic stroma is composed of cellular as well as extracellular components. Relatively little is known about the distribution of the different stromal components in the bone marrow. The knowledge on the bone marrow microenvironment is mainly based on studies in which in vitro long-term bone marrow cultures have been used. Although this culture system offers a unique possibility to study haematopoietic in vitro, it does not fully represent the complexity of intact bone marrow.

In the present study we describe the immunohistochemical distribution of different cellular and extracellular stromal components in frozen sections of rat bone marrow as well as in long-term bone marrow cultures, in order to compare the haematopoietic microenvironment used in in vitro studies in the in situ situation. We found that in situ a specific compartmentalization of stromal components exists in the bone marrow. Under culture conditions however, most stromal components are indeed present but the architecture present in the in situ situation had almost completely disappeared. The interaction between the different stromal elements was studied in the long-term bone marrow cultures. It appeared that under the chosen conditions, nodules were formed with a core of reticular cells and extracellular matrix. In close contact with this core immature macrophages appeared to proliferate and differentiate into mature, non-dividing macrophages.

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References

  • Barbé E, Damoiseaux JGMC, Döpp E et al. (1991) Characterization and expression of the antigen present on resident rat macrophages recognized by monoclonal antibody ED2. Immunobiology 1982:88–99

    Google Scholar 

  • Beelen RHJ, Eestermans IL, Döpp EA et al. (1987) Monoclonal antibodies ED1, ED2 and ED3 against rat macrophages: expression of recognized antigens in different stages if differentiation. Transplant Proc 19:3166–3170

    Google Scholar 

  • Ben-Ishay Z, Yoffey JM (1971) Reticular cells of erythroid islets of rat bone marrow in hypoxia and rebound, Reticuloendothel Soc 10:482–494

    Google Scholar 

  • Bentley SA (1984) The role and composition of the adherent layer in long-term bone marrow culture. In: Wright TN, and Greenberger JS (eds) Long term bone marrow culture. Alan R. Liss, New York, pp. 141–156

    Google Scholar 

  • Bessis M (1973) Erythroblastic islets. In: Bessis M (ed) Living blood cells and their ultrastructure. Springer-Verlag, Berlin, Heidelberg, New York

    Google Scholar 

  • Burtone MS (1958) Histochemical comparison of naphtholphosphate for demonstration of phosphatases. J Natl Cancer Inst 20:601–611

    Google Scholar 

  • Campbell A, Wicha MS, Long M (1985) Extracellular matrix promotes the growth and differentiation of murine haematopoestic cells in vitro. J Clin Invest 75:2085–2090

    Google Scholar 

  • Crocker PR, Morris L, Gordon S (1988) Novel cell surface receptors involved in interactions between stromal macrophages and haematopoietic cells. J Cell Sci Suppl 9:185–206

    Google Scholar 

  • Damoiseaux JGMC, Döpp EA, Neefjes JJ et al. (1989a) Heterogeneity of macrophages in the rat evidence by variability in determinants: two new antibodies against a heterodimer of 160 and 95 kD (CD11/CD18). J Leukocyte Biol 46:556–564

    Google Scholar 

  • Damoiseaux JGMC, Döpp EA, Beelen RHJ et al. (1989b) Rat bone marrow and monocyte cultures: influence of culture time and lymphokines on the expression of macrophage differentiation antigens. J Leukocyte Biol 46:246–253

    Google Scholar 

  • Dexter TM, Allen TD, Lajtha LG (1977) Conditions controlling the proliferation of hemmopoietic stem cells in vitro. J Cell Physiol 91:335–344

    Google Scholar 

  • Dijkstra CD, Damoiseaux JGMC (1993) Macrophage heterogeneity established by immunohistochemistry. In: Graumann W (ed) Progress in histochemistry and cytochemistry. Gustav Fisher Verlag, Stuttgart-New York, pp. 27–29

    Google Scholar 

  • Dorshkind K (1990) Regulation of hemopoiesis by bone marrow stromal cells and their products. Annu Rev Immunol 8:111–131

    Google Scholar 

  • Gartner S, Kaplan HS (1980) Long-term culture of human bone marrow cells. Proc Natl Acad Sci USA 77:4756–4759

    Google Scholar 

  • Gordon MY (1988) Annotation: extra cellular matrix of the marrow environment. Br J Haematol 70:1–5

    Google Scholar 

  • Hermans MHA, Opstelten D (1991) In situ visualization of hemopoietic cell subsets and stromal elements in rat and mouse bone marrow by immunostaining of frozen sections. J Histochem Cytochem 12:1627–1632

    Google Scholar 

  • Kaushansky K, Lin N, Adamson JW (1988) Interleukin 1 stimulates fibroblasts to synthesize granulocyte-macrophage and granulocyte colony-stimulating factors. Mechanisms of the hematopoietic response to inflammation. J Clin Invest 81:92–97

    Google Scholar 

  • Lombart Y, Ulrich B, Poindron P (1985) In vitro multiplication and apparently indefinite subcultures of normal mouse resident peritoneal macrophages. Biol Cell 53:219–230

    Google Scholar 

  • Morri M, Sadahira Y, Kawasaki S et al. (1990) Macrophage heterogeneity in bone marrow culture in vitro. J Cell Sci 95:481–482

    Google Scholar 

  • Morris L, Cocker PR, Fraser I et al. (1989) Expression of a divalent cation-dependent erythroblast adhesion receptor by stromal macrophages from murine bone marrow. J Cell Sci 99:141–147

    Google Scholar 

  • Rich IN (1986) A role for the macrophage in normal hemopoiesis. I. Functional capacity of bone-marrow derived macrophages to release hemopoietic growth factors. Exp Hematol 14:738–745

    Google Scholar 

  • Serke S, Sauberlich S, Abe Y et al. (1991) Analysis of CD34 positive haematopoietic progenitor cells from normal human adult peripheral blood. Flow cytometrical studies and in vitro colony (CFU-GM, BFU-E) assay. Ann Hematol 62:45–53

    Google Scholar 

  • Van den Berg TK, Döpp EA, Brevé JJP et al. (1989) The heterogeneity of the reticulum of rat peripheral lymphoid organs identified by monoclonal antibodies. Eur J Immunol 19:1747–1756

    Google Scholar 

  • Van den Berg TK, Brevé JJP, Damoiseaux JGMC et al. (1992) Sialoadhesin on macrophages: its identification as a lymphocyte adhesion molecule. J Exp Med 176:647–655

    Google Scholar 

  • Weiss L (1976) The hematopoietic microenvironment of the bone marrow and ultrastructural study of the stroma in rats. Anat Rec 1986:161–184

    Google Scholar 

  • Weiss L, Sakai H (1984) The hemopoietic stroma. Am J Anat 170:447–463

    Google Scholar 

  • Westermann J, Ronneberg S, Fritz FJ et al. (1989) Proliferation of macrophage subpopulations in the adult rat: comparison of various lymphoid organs. J Leukocyte Biol 46:236–247

    Google Scholar 

  • Whitelock CA, Witt ON (1982) Long-term culture of B lymphocytes and their precursors from murine bone marrow. Proc Natl Acad Sci USA 79:3608–3612

    Google Scholar 

  • Zuckermann KS, Wicha MS (1983) Extra cellular matrix production by the adherent cells of long term murine bone marrow cultures. Blood 61:540–547

    Google Scholar 

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Barbé, E., Damoiseaux, J.G.M.C., Döpp, E.A. et al. Stromal components in rat bone marrow frozen sections compared to long-term rat bone marrow cultures. Comparative Haematology International 5, 69–78 (1995). https://doi.org/10.1007/BF00638922

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