An in vitro system to screen for diarrheagenic chemicals
An in vitro system to screen for diarrheagenic chemicals
复制标题
DOI：
10.1007/bf00755142
复制
发表时间：
2004
影响因子：
6.1
通讯作者：
Y. Oshiro;J. Kachur;P. Balwierz;S. Won-Kim
中科院分区：
文献类型：
--
作者：
Y. Oshiro;J. Kachur;P. Balwierz;S. Won-Kim
我们检查了一个体外系统，使用已建立的肠道细胞系（T84人结肠癌）筛选致腹泻化学物质。细胞在 Millicell-PCF（聚碳酸酯膜）孔上生长。将细胞以约 5 × 106 个细胞/30mm 孔接种，并在 37°C 水饱和的 5% CO2 培养箱中培养 9-11 天。培养基是Ham's F12和Dulbecco's MEM的1:1混合物，含有5%胎牛血清和25pg/ml硫酸庆大霉素。将含有细胞的孔从培养箱中取出并安装在改进的尤斯室中以测量短路电流（Isc）。化学引起的 Psc 增加通常表明上皮细胞 Cl- 分泌，这与动物和人类的腹泻作用有关。在基底外侧添加胆碱能（毒蕈碱）激动剂卡巴胆碱、前列腺素 E2、16,16-二甲基前列腺素 E2 和毛喉素后，生长在 Millicell-PCF 膜上的 T84 细胞的 Isc 增加，而非致泻性前列腺素 D2 不影响 Isc。根据我们的结果，该体外系统有潜力适用于检测致腹泻化学物质的快速筛选。
We examined an in vitro system to screen for diarrheagenic chemicals using an established intestinal cell line (T84human colonic carcinoma). The cells were grown on Millicell-PCF (polycarbonate membrane) wells. The cells were seeded at approximately 5 × 106cells/30mm well and incubated for 9–11 days in a 5% C02incubator saturated with water at 37°C. The culture medium was a 1:1 mixture of Ham's F12 and Dulbecco's MEM with 5% fetal bovine serum and 25 pglml gentamicin sulfate. The well containing cells was removed from the incubator and mounted in a modified Ussing chamber for measurement of shortcircuit current (Isc). Chemical-induced increases in Psc are usually indicative of electrogenic epithelial Cl−secretion, which is associated with diarrheagenic effects in animals and humans. T84cells grown on Millicell-PCF membrane responded with an increase in Isc after basolateral addition of the cholinergic (muscarznic) agonist carbachol, prostaglandin E2, 16,16-dimethylprostaglandin E2, and forskolin, while non-diarrheagenic prostaglandin D2did not affect Isc. Based on our results, this in vitro system has the potential to be adapted as a rapid screen for detecting diarrheagenic chemicals.