Konsoula Roula, Barile Frank A
College of Pharmacy and Allied Health Professions, Department of Pharmaceutical Sciences, St. John's University, 8000 Utopia Parkway, Jamaica, NY 11439, USA.
J Pharmacol Toxicol Methods. 2007 Mar-Apr;55(2):176-83. doi: 10.1016/j.vascn.2006.06.001. Epub 2006 Jun 16.
The rat small intestinal cell line, IEC-18, was used as an in vitro model to differentiate between acute cytotoxicity (AC) and paracellular permeability (PP) of selected chemicals.
This study compares the low resistance rat intestinal mortal cell line, IEC-18 (transepithelial electrical resistance, TEER=160+/-10 Omega cm(2)) with the high resistance human intestinal cell line, Caco-2 (TEER=900+/-100 Omega cm(2)). The two cell lines differ in state of differentiation, TEER and paracellular permeability characteristics. The IEC-18 cell line is originated from the ileum and resembles more closely the small intestine than the Caco-2. Cytotoxicity was carried out using MTT cell viability assay in 96-well plates for 24-h exposure time. PP was measured using TEER (membrane integrity indicator) and PP markers such as [(3)H]-D-mannitol, lucifer yellow (LY) and FITC-dextran (fluorescein-dextran) on cells grown on inserts.
The data showed that there is a high correlation (R(2)=0.99) between MTT and TEER using IEC-18 cell for 24-h exposure time. IEC-18 is as sensitive as Caco-2 for both MTT and TEER measurements. Decrease in TEER is inversely proportional with increase in PP of tight junction indicators. There is a good correlation between IC(50)'s MTT, TEER and Registry of Cytotoxicity (RC) data.
Based on the results from the experiments, IEC-18 can be used as an in vitro model to differentiate between concentrations needed for AC and those required for PP.
大鼠小肠细胞系IEC-18被用作体外模型，以区分所选化学物质的急性细胞毒性（AC）和细胞旁通透性（PP）。
本研究将低电阻大鼠肠道永生细胞系IEC-18（跨上皮电阻，TEER = 160±10Ω·cm²）与高电阻人肠道细胞系Caco-2（TEER = 900±100Ω·cm²）进行比较。这两种细胞系在分化状态、TEER和细胞旁通透性特征方面存在差异。IEC-18细胞系起源于回肠，比Caco-2更接近小肠。细胞毒性通过在96孔板中使用MTT细胞活力测定法进行24小时暴露来检测。PP使用TEER（膜完整性指标）以及[³H]-D-甘露醇、荧光素黄（LY）和异硫氰酸荧光素-葡聚糖（荧光素-葡聚糖）等PP标志物在培养于小室中的细胞上进行测量。
数据显示，使用IEC-18细胞进行24小时暴露时，MTT与TEER之间存在高度相关性（R² = 0.99）。对于MTT和TEER测量，IEC-18与Caco-2一样敏感。TEER的降低与紧密连接指标的PP增加成反比。MTT的IC₅₀、TEER和细胞毒性登记册（RC）数据之间存在良好的相关性。
基于实验结果，IEC-18可作为体外模型，用于区分AC所需的浓度和PP所需的浓度。
