An improved procedure for the development of human mast cells from dispersed fetal liver cells in serum-free culture medium.
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, approximately 2 times the percentages in serum-containing media. By 6 weeks the percentages of mast cells were > or =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.
Kambe
N
N
Department of Internal Medicine, and Paediatrics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0263, USA.
Kambe
M
M
Chang
H W
HW
Matsui
A
A
Min
H K
HK
Hussein
M
M
Oskerizian
C A
CA
Kochan
J
J
Irani
A A
AA
Schwartz
L B
LB
D002452:Cell Count; D002454:Cell Differentiation; D002469:Cell Separation; D016895:Culture Media, Serum-Free; D046508:Culture Techniques / Q000379:methods*; D005333:Fetus / Q000166:cytology*; D006801:Humans; D008099:Liver / Q000166:cytology*; D008407:Mast Cells / Q000166:cytology*
D016895:Culture Media, Serum-Free