Isolation and Characterization of Human Intestinal Mast Cells
Mast cells are granulated immune cells typically located at barrier sites of the body, such as the skin and the mucosa of the respiratory, urogenital, and gastrointestinal tract. They are well known for their capacity to participate in the orchestration of inflammatory and immune responses by releasing a broad array of mediators as a consequence of IgE-dependent and IgE-independent activation. 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 visible 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 complicated. This chapter describes methods to isolate, purify, and culture mast cells from the human intestinal mucosa. Human mucosal mast cells can be used to characterize their mediators and to study the mechanisms of human mast cell activation, signal transduction, and exocytosis in response to specific stimuli.
Key wordsMast cells Human Intestinal Gut Bowel Cell isolation Cell culture Cell activation Mediator release assay
The authors thank all former and current colleagues and in particular C. A. Dahinden, K. Wordelmann, S. Schwengberg, C. T. Mierke, G. Weier, T. Gebhardt, L. E. Sander, S. P. Frank, A. Mrasori, K. Feuser, and Y. Soltow who were involved in establishing the methods described here.
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