Purification of human colonic and gastric mast cells
| Autor: | P. M. Shah, Henrik Vendelbo Nielsen, T. Engberg Damsgaard, Peter Oluf Schiøtz, Steffen Husby |
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| Rok vydání: | 1998 |
| Předmět: |
Adult
Pathology medicine.medical_specialty Cell Survival Colon Immunology Cell Separation Biology Immunoglobulin E Histamine Release Dynabeads chemistry.chemical_compound Concanavalin A Gastric mucosa medicine Humans Immunology and Allergy Collagenases Mast Cells Intestinal Mucosa Interleukin 5 Calcimycin Aged Ionophores CD117 Stomach Middle Aged Mast cell Molecular biology Interleukin 33 medicine.anatomical_structure chemistry Gastric Mucosa biology.protein Histamine |
| Zdroj: | Journal of Immunological Methods. 214:141-148 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/s0022-1759(98)00044-1 |
| Popis: | We have developed a method to purify mast cells from enzymatic isolates of human colonic mucosa (HCM) and submucosa/muscle (HCS), and gastric mucosa (HGM) and submucosa/muscle (HGS). The purification of mast cells from these enzymatic isolates involves positive affinity-magnetic selection of mast cells using a monoclonal antibody specific for the c-kit receptor tyrosine kinase (CD117). The monoclonal antibody is coupled to Dynabeads for positive affinity selection of c-kit receptor positive cells which includes mast cells. This selection procedure generates preparations of mast cells from HCM, HCS, HGM and HGS that are 80% pure. The purified mast cells were microscopically normal and viable (>85%). The functionality of purified mast cells was examined by studying the effect of anti-human IgE, Concanavalin A (Con A) and calcium ionophore A23187 on histamine release. These results show that this purification procedure generates microscopically normal, viable and functional mast cells. This method of purifying human gastrointestinal tissue mast cells may be a valuable tool for the further study of mast cell heterogeneity and the role of mast cells in the gastrointestinal tract. |
| Databáze: | OpenAIRE |
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