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After washing the cells twice with PBS, they were grown in IMDM media (Invitrogen) supplemented with 20% horse serum (Omega Scientific), 1X P/S and 10% BHK/HM-5 conditioned medium as a source of GM-CSF, for 9 days without splitting cells.
Recent data from murine studies have identified Th17 cells as a key source of GM-CSF and suggest that T cell derived GM-CSF is instrumental in the induction of autoimmune disease.
Terminal granulocytic differentiation was induced by removing IL-3 and SCF and adding 10% BHK-HM5 CM as a source of granulocyte/macrophage-colony stimulating factor (GM-CSF).
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