Stable Heparin-Producing Cell Lines Derived from the Furth Murine Mastocytoma
Glycosaminoglycans/metabolism, Heparin/biosynthesis*, Mast cells/cytology
Heparin, Mast cells, Glycosaminoglycans
Biology | Ecology and Evolutionary Biology
Stable cell lines that synthesize heparin have been established from the Furth murine mastocytoma. The parental line(MST)divides in suspension every 14-18h in growth medium supplemented with fetal bovine serum or defined growth factors. Adherent subclones were selected by adhesion to plastic culture vessels. Both adherent and nonadherent cells contain about 0.4 µg of glycosaminoglycan hexuronic acid per 106 cells, composed of 80% heparin and 20% chondroitin sulfate E. Deaminative cleavage of MST heparin by HNO2 at pH 1.5 released disaccharides that were similar in composition to those obtained from commercial heparin, except that disaccharides containing
3,6-0-disulfated GlcN units were not found. Greater than 90% of the glycosaminoglycans were stored in cytoplasmic granules, and challenge of the cells with dinitrophenylated bovine serum albumin and anti-dinitrophenyl IgE released a portion of the stored material. Growth studies of subclones showed that MST cells tolerate a 10-fold variation in glycosaminoglycan content. Incubation of cells with sodium chlorate reduced glycosaminoglycan sulfation by >95% without affecting cell growth. Thus, granule glycosaminoglycans appear to be nonessential for growth of MST cells.
Montgomery, Rebecca I.; Lidholt, Kerstin; Flay, Nina; Liang, Jonathan; Vertel, Barbara; Lindahl, Ulf; and Esko, Jeffrey D., "Stable Heparin-Producing Cell Lines Derived from the Furth Murine Mastocytoma" (1992). Biology and Environmental Science Faculty Publications. 40.
Montgomery, Rebecca I., Lidholt, Kerstin, Flay, Nina W., Liang, Jonathan, Vertel, Barbara, Lindahl, Ulf, Esko, Jeffrey D. Stable heparin-producing cell lines derived from the Furth murine mastocytoma. PNAS:89, 11327-11331, December 1992.