Intestinal absorption of pesticides across Caco-2 cell monolayers

ABSTRACT: A monolayer of Caco-2 cells was cultured in transwells, maintaining a stable TEER for at least one week. On the apical side, cells were exposed to pesticides in HBSS (without additives) for a maximum of 5 hours. Samples were collected over time from both the basolateral and apical compartments: either 50 µL or 100 µL immediately after exposure (time point 0.01), and at 30 minutes, 1 hour, 2 hours, and 5 hours. Each sample was diluted with an equal volume (50 µL or 100 µL) of LC-MS/MS grade methanol (cyclohexane for endosulfan). Pesticide concentrations were quantified by LC-MS/MS and used to determine apical-to-basolateral transport (absorption) across the intestinal epithelial barrier. In parallel, the cytotoxicity of the pesticides towards Caco-2 cells was assessed with an alamarBlue (resazurin) mitochondrial-activity assay, to confirm that the tested concentrations did not compromise monolayer viability. The Transwell permeability procedure was adapted from Hubatsch et al., Nature Protocols 2007 (doi:10.1038/nprot.2007.303).

ORGANISM(S): Homo sapiens

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PROVIDER: S-VHPS18 | biostudies-other |

REPOSITORIES: biostudies-other