Stable heparin-producing cell lines derived from the Furth murine mastocytoma.

Abstract

(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) ,uCi/ml; 40 Ci/mmol) (both from New England Nuclear).Radioactive GAGs were isolated by anion-exchange chromatography and ethanol precipitation as described by Bame and Esko (20), except that samples bound to DEAE-Sephacel (Pharmacia LKB) were washed with 0.25 M NaCl in 20 mM sodium acetate (pH 6.0) and eluted with 2.0 M NaCl in acetate buffer. Samples were 3-eliminated with 0.5 M NaOH for 24 h at 40C and reduced with 1 M NaBH4 (21) to remove any covalently linked peptides. Chondroitin sulfate was depolymerized with chondroitinase ABC according to the manufacturer's directions (ICN), and the enzyme was inactivated by boiling.To measure GAGs chemically, preparations were first treated with 10 units of DNase I and RNase A (Sigma) at 10 ,ug/ml to degrade nucleic acids. The GAGs were repurified by anion-exchange chromatography, and uronic acid was

Mouse Mastocytoma Cells Synthesize Undersulfated Heparin and Chondroitin Sulfate in the Presence of Brefeldin A

“…The low level of 3-O-sulfation may be due to the transformed state of the cells. Also in established cell lines derived from this tumor, the heparin synthesized contains much less 3-O-sulfated products than commercial heparin (25 a 3 H-Labeled disaccharides obtained by N-deacetylation followed by deamination at pH 3.9 and 1.5 (resulting in cleavage of all glucosaminidic linkages) and reduction with NaBH 4 , were analyzed by anionexchange HPLC as described under "Experimental Procedures." b ND, not detected.…”

Cloning, Golgi Localization, and Enzyme Activity of the Full-length Heparin/Heparan Sulfate-Glucuronic Acid C5-epimerase

“…MST cells were derived from the Furth murine mastocytoma (10). CHO cells were grown in Ham's F-12 medium (Life Technologies, Inc.), and MST cells were grown in RPMI 1640 medium.…”

Distinct role for c-kit receptor tyrosine kinase and SgIGSF adhesion molecule in attachment of mast cells to fibroblasts

“…Two mastocytoma cell lines, FMA/3 and P-815, 33,34 were grown in a-MEM supplemented with 10% FCS. Another mastocytoma cell line, MST, 35 was kindly provided by Dr JD Esko (University of California, San Diego, CA, USA), and grown in RPMI 1640 (Sigma Chemical Co., St Louis, MO, USA) supplemented with 10% FCS. NIH/3T3 fibroblasts and C2 retrovirus packaging cells were maintained in DMEM (ICN Biomedicals) supplemented with 10% FCS.…”