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An improved procedure for the development of human mast cells from dispersed fetal liver cells in serum-free culture medium
Authors:Kambe N  Kambe M  Chang H W  Matsui A  Min H K  Hussein M  Oskerizian C A  Kochan J  Irani A A  Schwartz L B
Affiliation:

a Department of Internal Medicine, and Paediatrics, Medical College of Virginia, Virginia Commonwealth University, P.O. Box 980263, Richmond, VA 23298-0263, USA

b Hoffmann-La Roche, Nutley, NJ, USA

Abstract:The in vitro development of human mast cells from fetal liver cells with recombinant human stem cell factor in serum-containing RPMI was compared to that in AIM-V media with and without serum. Compared to serum-containing media, AIM-V medium caused mast cells to develop earlier and in greater numbers. By 2 weeks, about 60% of cells in serum-free AIM-V medium were phenotypic mast cells, 2 times the percentages in serum-containing media. By 6 weeks the percentages of mast cells were ≥80% under all conditions, but the number of mast cells was 3–4-fold greater in serum-free AIM-V medium than in serum-supplemented media. Mast cells obtained in serum-free AIM-V medium exhibited rounded nuclei, like tissue-derived mast cells; mast cells obtained in serum-supplemented media had segmented nuclei. By 10–12 weeks of culture about 40% of the AIM-V-derived cells showed strong chymase immunocytochemical staining, a pattern observed for only 14% of the cells in serum-containing media. AIM-V medium is a suitable medium for the development of human mast cells in vitro, and permits an earlier, more selective and greater expansion of mast cells than serum-containing media.
Keywords:Human mast cell   Serum-free culture medium   Tryptase   Chymase   Kit
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