[The culture of murine fetal skin-derived mast cells]

Objective To isolate, induce and culture murine fetal skin-derived mast cells. Methods Day 14-16 murine fetal skin cells were separated and cultured in the presence of recombinant mouse interleukin-3 (rmIL-3) and recombinant mouse stem cell factor (rmSCF) to induce the maturation of mast cells. After cultured for 2 weeks, the mast cells were obtained by gradient centrifugation. The cells were stained with toluidin blue or anti-mast cell tryptase antibody to identify the purity. FcepsilonR I and CD117, which were the surface markers of mature mast cells, were identified with flow cytometry. Results After cultured for 14 days and purified, the cytoplasm was observed filled with typical granules using toluidin blue staining or with tryptase positive substance using immunocytochemistry. The expression of FcepsilonRIand CD117 was found in 97% cells. Conclusion Murine fetal skin-derived mast cells were cultured successfully and a large number of highly purified mast cells were obtained.