Murine basophil—mast differentiation: Toward optimal conditions for selective growth and maturation of basophil—mast or allied cells

Abstract

Recent investigations revealed that basophil-mast cells were related to the hemopoietic system. Strikingly, murine bone marrow showed a singular paucity in cells with basophil-mast features; moreover in clonogenic assays (methylcellulose, agarose) bone marrow was found to be manifestly poor in basophil-mast progenitor cells. Our work brought to light several new facts concerning the culture and differentiation of this cell type: 1 degree pure and mixed mast clones can be derived in large numbers from bone marrow, provided progenitors are cultured in collagen matrix. Up to 1,382 hemopoietic clones were analysed in situ after staining: 30% contained mast cells (34 per 10(5) cells), thus the basophil-mast lineage was one of the most frequent. We concluded that other cloning media were noticeably nonoptimal for the growth and/or maturation of mast cells. We suggested that collagen and the molecular edifices derived from it, both found in variable amounts in the natural mast environments, should play essential roles in mast phenotype expression. 2 Degrees cholera toxin (CT) selectively eradicated nonmast progenies: mast progenitors and mast progenies were resistant. In this way, pure and rapidly expanding mast cell clones were obtained at a frequency never reported before. CT possibly acts both directly, as a stimulator of mast cell proliferation, or indirectly on marrow subpopulations which repress basophil-mast cell growth and maturation. In vitro culture conditions, specifically designed for basophil-mast lineage, should prove of interest in the search for an unifying hypothesis concerning the multiple forms of mast cells found in various tissues.

Establishment and characterization of continuous hematopoietic progenitors-derived pig normal mast cell lines

“…These MC express relatively low amounts of intracellular histamine as compared to their connective tissue-derived counterparts, poorly sulfated proteoglycans and low amounts of membrane FcqRI, the expression of which being up-regulated by several days treatment with monomeric IgE (Kawakami and Galli, 2002). Moreover, and contrasting to what is observed with our PMC lines, the growth of BMMC is limited in time, the cells undergoing apoptosis after a few months in culture, despite the continuous presence of murine IL-3 (Lanotte et al, 1986). In addition, and contrasting again with what we have observed with our continuous PMC lines, BMMC cannot be preserved by freezing with conventional, DMSO-containing medium.…”