41] Murine mast cell growth factor

Abstract

This chapter discusses the murine mast cell growth factor. Murine MCGF is a trypsin and neuraminidase-sensitive glycoprotein 3 present in mitogen stimulated splenocyte conditioned medium with the capacity to induce and maintain the continued proliferation of murine mast cells from their marrow derived precursors. MCGF purified from spleen conditioned medium 2 has a molecular weight of approximately 35,000 and is devoid of IL-2 and CSF activities. Mast cell lines are generated from Dexter-type long-term bone marrow cultures (LMC). Mast cell lines are then established from LMC. Cells harvested from LMC of BDF1 mice are transferred to secondary cultures in RPMI 1640 medium containing 10% FCS and 10% WEHI-CM. Established pure mast cell lines are maintained by passaging twice a week under the same culture conditions. Fresh bone marrow cells may also be used to establish mast cell lines. Prior to use in MCGF assay, the mast cells are washed with RPMI 1640 medium containing 10% FCS and resuspended at 105 cells/ml. The chapter further describes the preparation of WEHI-CM.