Roula Konsoula1, Frank A Barile. 1. College of Pharmacy and Allied Health Professions, Department of Pharmaceutical Sciences, St. John's University, 8000 Utopia Parkway, Jamaica, NY 11439, USA.
Abstract
INTRODUCTION: 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. METHODS: 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. RESULTS: 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. DISCUSSION: 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.
INTRODUCTION: 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. METHODS: 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. RESULTS: 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. DISCUSSION: 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.
Authors: Sotirios I Sinis; Chrissi Hatzoglou; Konstantinos I Gourgoulianis; Sotirios G Zarogiannis Journal: Front Physiol Date: 2018-03-29 Impact factor: 4.566
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