The effects of recombinant ovine interleukin-3 and recombinant ovine stem cell factor on the growth and mediator expression of caprine and ovine bone marrow-derived mast cells.
The growth of ovine and caprine mast cells in bone marrow cultures has been achieved using recombinant ovine interleukin-3 (rOvIL-3) and recombinant ovine stem cell factor (rOvSCF). After approximately 2-3 weeks' growth in optimal concentrations of either rOvIL-3 alone or a combination of rOvIL-3 and rOvSCF, the majority of the cells produced in bone marrow culture from both species were mast cells. The significant increase in the total numbers of cells and survival times of the cultures when both cytokines were present compared to either alone, indicated synergy between rOvIL-3 and rOvSCF on mast cell growth. Ovine and caprine cells cultured in rOvIL-3 alone produced a four-fold increase in cell numbers compared with medium only controls. The resulting cultures contained up to 52% mast cells by day 18 and had a lifespan of 3-4 weeks. In contrast, cells from both species grown in both rOvIL-3 and rOvSCF produced up to six times more cells than the equivalent rOvIL-3 stimulated cultures, contained up to 69% mast cells by day 21 and could be maintained for at least 6 weeks. Ovine cells grown in rOvIL-3 alone or rOvIL-3 and rOvSCF contained significantly more aryl-sulfatase and serine protease but similar amounts of beta-hexosaminidase compared with caprine cells during the second week of culture. There were no significant differences in the granule-associated mediator content of cells from either individual species grown in rOvIL-3 alone compared with those grown in rOvIL-3 and rOvSCF during the first 21 days of culture.
Macaldowie
C N
CN
Moredun Research Institute, Edinburgh, UK.
Huntley
J F
JF
Mackellar
A
A
McInnes
C
C
Haig
D M
DM
D000818:Animals; D001192:Arylsulfatases / Q000378:metabolism; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000201:enzymology; D002455:Cell Division / Q000187:drug effects; D053818:Chymases; D004305:Dose-Response Relationship, Drug; D006041:Goats; D007377:Interleukin-3 / Q000494:pharmacology*; D008407:Mast Cells / Q000187:drug effects* / Q000201:enzymology; D011994:Recombinant Proteins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000378:metabolism; D012756:Sheep; D019089:Stem Cell Factor / Q000494:pharmacology*; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D007377:Interleukin-3; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D001192:Arylsulfatases; D001619:beta-N-Acetylhexosaminidases; D012697:Serine Endopeptidases; D053818:Chymases