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[Purification and reactivity of human lung mast cells]
K Takahashi1, R Soda, T Kishimoto
1Second Department of Internal Medicine, Okayama University Medical School.
Abstract:
To clarify the pathogenesis of allergic pulmonary diseases by analysis of the reactivity of human lung mast cells, we established a method of dispersion and purification of mast cells from human lung tissue. This method consisted of 4 steps; 1) mincing by scissors, 2) enzymatic treatment by a pronase-chymopapain and collagenase-elastase mixture, 3) percoll centrifugation and 4) exclusion of adherent cells. Using this method, dispersed human lung mast cells were obtained with 38.8% purity and more than 95% viability. These mast cells contained 4.1 pg of histamine per cell, which showed these cells had mild spontaneous histamine release after treatment. The mast cells released histamine in a dose-dependent manner after treatment with calcium ionophore A 23187 and anti-IgE, and these phenomena were dependent on extracellular calcium ions. However, the cells did not release histamine with less than 100 micrograms/ml of compound 48/80. These results indicate that the human lung mast cells obtained by this method are useful to make immunological and pharmacological analyses in allergic lung diseases.