Stable heparin-producing cell lines derived from the Furth murine mastocytoma.
Stable heparin-producing cell lines derived from the Furth murine mastocytoma.
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源自 Furth 鼠肥大细胞瘤的稳定产生肝素的细胞系。
DOI：
10.1073/pnas.89.23.11327
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发表时间：
1992
影响因子：
11.1
通讯作者：
Esko,JD
中科院分区：
文献类型：
--
作者：
Montgomery,RI;Lidholt,K;Flay,NW;Liang,J;Vertel,B;Lindahl,U;Esko,JD
合成肝素的稳定细胞系已经从Furth鼠肥大细胞瘤中建立。亲本系（MST）在补充有胎牛血清或确定的生长因子的生长培养基中每14-18小时在悬浮液中分裂。通过粘附到塑料培养容器上来选择粘附的亚克隆。贴壁和非贴壁细胞每10（6）个细胞含有约0.4微克的糖胺聚糖己糖醛酸，由80%肝素和20%硫酸软骨素E组成。在pH1.5时用HNO 2将MST肝素脱氨基裂解，释放出的二糖与从商业肝素中获得的二糖在组成上相似，只是没有发现含有3，6-O-取代GlcN单元的二糖。用二硝基苯基化牛血清白蛋白和抗二硝基苯基IgE攻击细胞释放了一部分储存的物质。亚克隆的生长研究表明，MST细胞耐受10倍的糖胺聚糖含量的变化。用氯酸钠孵育细胞使糖胺聚糖硫酸化减少> 95%，而不影响细胞生长。因此，颗粒糖胺聚糖似乎是MST细胞生长的非必需的。
Stable cell lines that synthesize heparin have been established from the Furth murine mastocytoma. The parental line (MST) divides in suspension every 14-18 h 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 micrograms of glycosaminoglycan hexuronic acid per 10(6) 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-O-desulfated 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.