Abstract
Mast cells are bone-marrow-derived tissue cells typically located at barrier sites of the body, such as skin, mucosal barriers, or blood barriers, that is, around blood vessels. This location suggests that mast cells might have a function as immunological “gatekeepers” or “watch dogs” and, indeed, some recent functional data support this idea. Mast cells derive from myeloid progenitors, but in contrast to other myeloid cells, they leave the bone marrow in an immature state; therefore, mast cells are not found in the blood under normal conditions. For full maturation, the tissue environment is necessary. Thus, mature mast cells can be only isolated from tissue such as skin or mucosal sites, which makes mast cell isolation rather complicated. Alternatively, mast cell progenitors can be isolated from the bone marrow, peripheral blood, or cord blood, which is easier but requires subsequent in vitro maturation of mast cells as far as possible using cytokines. This chapter describes a rather new technique of mast cell isolation from human intestinal mucosal tissue yielding approx 1–5 million pure and viable human mast cells suitable to perform functional and cell culture experiments.
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References
Weidner, N. and Austen, K. F. (1990) Evidence for morphologic diversity of human mast cells. An ultrastructural study of mast cells from multiple body sites. Lab. Invest. 63, 63–72.
Metcalfe, D. D., Baram, D., and Mekori, Y. A. (1997) Mast cells. Physiol. Rev. 77, 1033–1079.
Bischoff, S. C., Wedemeyer, J., Herrmann, A., et al. (1996) Quantitative assessment of intestinal eosinophils and mast cells in inflammatory bowel disease. Histopathology 28, 1–13.
Bischoff, S. C. and Sellge, G. (2002) Mast cell hyperplasia: role of cytokines. Int. Arch. Allergy Immunol. 127, 118–122.
Gurish, M. F., Tao, H., Abonia, J. P., et al. (2001) Intestinal mast cell progenitors require CD49dbeta7 (alpha4beta7 integrin) for tissue-specific homing. J. Exp. Med. 194, 1243–1252.
Craig, S. S., Schechter, N. M., and Schwartz, L. B. (1989) Ultrastructural analysis of maturing human T and TC mast cells in situ. Lab. Invest. 60, 147–157.
Lorentz, A., Schwengberg, S., Mierke, C., Manns, M. P., and Bischoff, S. C. (1999) Human intestinal mast cells produce IL-5 in vitro upon IgE receptor crosslinking and in vivo in the course of intestinal inflammatory disease. Eur. J. Immunol. 29, 1496–1503.
Gelbmann, C. M., Mestermann, S., Gross, V., Kollinger, M., Scholmerich, J., and Falk, W. (1999) Strictures in Crohn’s disease are characterised by an accumulation of mast cells colocalised with laminin but not with fibronectin or vitronectin. Gut 45, 210–217.
Bischoff, S. C., Lorentz, A., Schwengberg, S., Weier, G., Raab, R., and Manns, M. P. (1999) Mast cells are an important cellular source of tumour necrosis factor alpha in human intestinal tissue. Gut 44, 643–652.
Bischoff, S. C., Schwengberg, S., Wordelmann, K., Weimann, A., Raab, R., and Manns, M. P. (1996) Effect of c-kit ligand, stem cell factor, on mediator release by human intestinal mast cells isolated from patients with inflammatory bowel disease and controls. Gut 38, 104–114.
Bischoff, S. C. and Dahinden, C. A. (1992) c-kit ligand: a unique potentiator of mediator release by human lung mast cells. J. Exp. Med. 175, 237–244.
Gibbs, B. F., Wierecky, J., Welker, P., Henz, B. M., Wolff, H. H., and Grabbe, J. (2001) Human skin mast cells rapidly release preformed and newly generated TNF-alpha and IL-8 following stimulation with anti-IgE and other secretagogues. Exp. Dermatol. 10, 312–320.
Lowman, M. A., Rees, P. H., Benyon, R. C., and Church, M. K. (1988) Human mast cell heterogeneity: histamine release from mast cells dispersed from skin, lung, adenoids, tonsils, and colon in response to IgE-dependent and nonimmunologic stimuli. J. Allergy Clin. Immunol. 81, 590–597.
Pawankar, R., Okuda, M., Yssel, H., Okumura, K., and Ra, C. (1997) Nasal mast cells in perennial allergic rhinitics exhibit increased expression of the Fc epsilonRI, CD40L, IL-4, and IL-13, and can induce IgE synthesis in B cells. J. Clin. Invest. 99, 1492–1499.
Bischoff, S. C., Sellge, G., Schwengberg, S., Lorentz, A., and Manns, M. P. (1999) Stem cell factor-dependent survival, proliferation and enhanced releasability of purified mature mast cells isolated from human intestinal tissue. Int. Arch. Allergy Immunol. 118, 104–107.
Bischoff, S. C., Sellge, G., Lorentz, A., Sebald, W., Raab, R., and Manns, M. P. (1999) IL-4 enhances proliferation and mediator release in mature human mast cells. Proc. Natl. Acad. Sci. USA 96, 8080–8085.
Schulman, E. S., Macglashan, D. W., Peters, S. P., Schleimer, R. P., Newball, H. H., and Lichtenstein, L. M. (1982) Human lung mast cells: purification and characterization. J. Immunol. 129, 2662–2667.
Befus, A. D., Dyck, N., Goodacre, R., and Bienenstock, J. (1987) Mast-cells from the human intestinal lamina propria—isolation, histochemical subtypes, and functional-characterization. J. Immunol. 138, 2604–2610.
Bischoff, S. C., Schwengberg, S., Raab, R., and Manns, M. P. (1997) Functional properties of human intestinal mast cells cultured in a new culture system: enhancement of IgE receptor-dependent mediator release and response to stem cell factor. J. Immunol. 159, 5560–5567.
Gebhardt, T., Sellge, G., Lorentz, A., Raab, R., Manns, M. P., and Bischoff, S. C. (2002) Cultured human intestinal mast cells express functional IL-3 receptors and respond to IL-3 by enhancing growth and IgE receptor-dependent mediator release Eur. J. Immunol. 32, 2308–2316.
Mierke, C. T., Ballmaier, M., Werner, U., Manns, M. P., Welte, K., and Bischoff, S. C. (2000) Human endothelial cells regulate survival and proliferation of human mast cells. J. Exp. Med. 192, 801–811.
Sellge, G., Lorentz, A., Gebhardt, T., et al. (2004) Human intestinal fibroblasts prevent apoptosis in human intestinal mast cells by a mechanism independent of stem cell factor, IL-3, IL-4, and nerve growth factor. J. Immunol. 172, 260–267.
Lorentz, A., Schwengberg, S., Sellge, G., Manns, M. P., and Bischoff, S. C. (2000) Human intestinal mast cells are capable of producing different cytokine profiles: role of IgE receptor cross-linking and IL-4. J. Immunol. 164, 43–48.
Bischoff, S. C., Schwengberg, S., Lorentz, A., et al. (2004) Substance P and other neuropeptides do not induce mediator release in isolated human intestinal mast cells. Neurogastroenterol. Motil. 16, 185–193.
Acknowledgments
The authors thank all former and current colleagues and in particular C. A. Dahinden, K. Wordelmann, S. Schwengberg, C. Mierke, A. Lorentz, G. Weier, N. Steegmann, T. Gebhardt, and A. Radke, who were involved in establishing the methods described here.
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Sellge, G., Bischoff, S.C. (2006). Isolation, Culture, and Characterization of Intestinal Mast Cells. In: Krishnaswamy, G., Chi, D.S. (eds) Mast Cells. Methods in Molecular Biology, vol 315. Humana Press. https://doi.org/10.1385/1-59259-967-2:123
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DOI: https://doi.org/10.1385/1-59259-967-2:123
Publisher Name: Humana Press
Print ISBN: 978-1-58829-374-9
Online ISBN: 978-1-59259-967-7
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