Isolation and study of functional mast cells from lung and mesentery of the guinea pig.
A method is described for the dissociation of guinea pig lung and mesentery into their component cells. The method comprises incubation of the tissues with the enzyme pronase in Ca++- and Mg++-free saline followed by mechanical dispersion and yields on average 3-8% free mast cells. These cells are morphologically intact and viable. They retain an active sensitization or can be passively sensitized. The levels of cAMP and cGMP in the whole cell suspension respond to catecholamine, histamine or cholinergic stimulation in a way similar to that observed in the corresponding undispersed tissue. Although the separation of guinea pig mast cells from other cells proved to be more difficult than the purification of rat mast cells, an inrichment of greater than 50% has been achieved. The mast cells isolated by this method proliferate in vitro and have been cultured successfully for several months.
D000707:Anaphylaxis; D000818:Animals; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D005260:Female; D006168:Guinea Pigs / Q000276:immunology; D006636:Histamine Release; D007116:Immunization, Passive; D008168:Lung / Q000166:cytology* / Q000276:immunology; D008297:Male; D008407:Mast Cells / Q000276:immunology / Q000502:physiology* / Q000648:ultrastructure; D008643:Mesentery / Q000166:cytology* / Q000276:immunology; D008854:Microscopy, Electron; D009928:Organ Specificity; D011402:Pronase
D000242:Cyclic AMP; D011402:Pronase; D006152:Cyclic GMP