Study on the culture of mature human mast cells in vitro

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To obtain a sufficient number of human mast cells by culturing mature mast cells in vitro, basic study was undertaken mainly using rat mast cells.1.Rat peritoneal mast cells were purified by Percoll density gradient centrifugation. Although IgE-dependent histamine release from purified mast cells was very low, reconstitution with separated cells recovered the lowered histamine release. Recovery of the histamine release was dependent upon the dose of separated cells added and the contact period of mast cells and separated cells. Incubation supernatant of separated cells failed to recover the histamine release from purified mast cells.2.Stem cell factor (SCF) could not induce histamine release from rat peritoneal mast cells, but potentiate the IgE-dependent histamine release. Although anti-SCF antibody neutralize the potentiation of histamine release by SCF,it did not affect the recovery of histamine release from purified mast cells by reconstitution with separated cells.3.Mastcytosis could be induced in the rat peritoneal cavity by IgE-antigen interaction. Prior priming with antigen was essential for the mastcytosis, suggesting that antigen-primed immunocompetent cells provide a support the mast cell increase.4.Peritoneal cells from immunized and sensitized rats were cultured in the presence of spleen cells from the same rats. Antigen stimulation caused an increase in the number of mast cell-like cells in 3-4 weeks.5.A method of dispersion of human and monkey lung tissue with collagenase and hyarulonidase was examined.Basic data obtained from experiments using rat mast cells will be applied to human mast cells.