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 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.
Allergy and Immunology Laboratory, Department of Paediatrics, Aarhus University Hospital, Denmark.
P M
PM
Shah
S
S
Husby
T E
TE
Damsgaard
H V
HV
Nielsen
P O
PO
Schiøtz
D000328:Adult; D000368:Aged; D000001:Calcimycin / Q000494:pharmacology; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D017364:Collagenases / Q000378:metabolism; D003106:Colon / Q000166:cytology*; D003208:Concanavalin A / Q000494:pharmacology; D005753:Gastric Mucosa / Q000166:cytology; D006636:Histamine Release / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology; D007476:Ionophores / Q000494:pharmacology; D008407:Mast Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D008875:Middle Aged; D013270:Stomach / Q000166:cytology*
D007476:Ionophores; D003208:Concanavalin A; D000001:Calcimycin; D017364:Collagenases