Purification of human colonic and gastric mast cells

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 ce...

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Bibliographic Details
Published in:Journal of immunological methods 1998-05, Vol.214 (1), p.141-148
Main Authors: Shah, P.M, Husby, S, Engberg Damsgaard, T, Nielsen, H.V, Schiøtz, P.O
Format: Article
Language:English
Subjects:
Adult
Aged
Ast cells
Biological and medical sciences
c-kit receptor
Calcimycin - pharmacology
CD117
Cell Separation - methods
Cell Survival - physiology
Collagenases - metabolism
Colon - cytology
Concanavalin A - pharmacology
Diverse techniques
Fundamental and applied biological sciences. Psychology
Gastric Mucosa - cytology
Gastrointestinal tract
Histamine Release - drug effects
Histamine Release - physiology
Humans
Intestinal Mucosa - cytology
Ionophores - pharmacology
Mast Cells - cytology
Mast Cells - drug effects
Mast Cells - physiology
Middle Aged
Molecular and cellular biology
Positive affinity-magnetic selection
Stomach - cytology
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Summary: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.
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(98)00044-1