Factors affecting the growth and maintenance of human skin mast cells in cell culture.
The isolation and kinetics of survival of human mast cells from newborn and adult skin is described. Recombinant human interleukins and conditioned medium from several human cell lines were tested for their ability to maintain mast cells in vitro. Growth medium supplemented with IL-2, IL-4 and conditioned medium from a mixed lymphocyte culture enhanced mast cell survival resulting in a 30-fold increase in survival (relative to that obtained with non-supplemented medium) at 7 days, and a 15-fold increase at 15 days. Cell survival for time periods longer than 21 days was not observed. Inclusion of cAMP, agents that elevate cAMP, insulin, and epidermal growth factor in supplemented growth medium prevented the enhanced survival by 40-70%. Incorporation of bromodeoxyuridine (BrdU) into mast cells in 3-day cultures demonstrated that 15% of the mast cell population was capable of proliferation. At 21 days, no incorporation of BrdU could be detected. After 3 days in culture mast cells released 16% of their histamine stores in response to A23187 and 10% in response to anti-human IgE. Electron microscopy of cultured cells at 3 days revealed cells with both intact and empty mast cell granules. These results demonstrate that human skin mast cells proliferate in response to cytokines and release histamine when stimulated with classical secretagogues. Since human skin mast cells retain these basic properties in vitro, they may be useful in further functional studies involving their proliferation and secretion.
Rein
G
G
Psoriasis Research Institute, Palo Alto, Calif.
Karasek
M A
MA
D000328:Adult; D002455:Cell Division; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D000242:Cyclic AMP / Q000494:pharmacology; D006636:Histamine Release; D006801:Humans; D007378:Interleukins / Q000494:pharmacology; D008407:Mast Cells / Q000166:cytology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D012867:Skin / Q000166:cytology*
D003470:Culture Media; D007378:Interleukins; D011994:Recombinant Proteins; D000242:Cyclic AMP