Long-term in vitro culture of murine mast cells. I. Description of a growth factor-dependent culture technique
Tertian, G.; Yung, Y.P.; Guy-Grand, D.; Moore, M.A.
Journal of Immunology 127(2): 788-794
1981
ISSN/ISBN: 0022-1767 PMID: 7019332 Document Number: 177495
A feeder layer independent long-term in vitro culture system for murine mast cells is described. Concanavalin A-activated murine splenic leukocyte-conditioned medium, prepared under conditions optimal for T cell growth factor production, contains a growth-promoting activity for murine mast cells identified by their morphology, characteristic ultrastructure of the granules, positive reactions with toluidine blue and alcian blue, presence of receptors for IgG and IgE, and presence of histamine, serotonin, L-dopa, 5-hydroxytryptophan and sulfated products within the cytoplasm. After 2-3 wk of culture in the presence of the conditioned medium, mast cell lines were established from various sources initially devoid of matured mast cells. Such sources included spleen and bone marrow of athymic nude mice, long-term cultured marrow cells and T cell-depleted normal marrow. Cultured mast cells are absolutely dependent upon the conditioned medium-derived growth factor(s) for growth and viability. Death ensues within 24 h in the absence of the factor(s). Established mast cell lines were maintained in exponential growth for over 1 yr by passaging in the conditioned medium every 3-7 days.