2008
DOI: 10.1016/j.tiv.2008.02.023
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Evaluation of in vitro cytotoxicity and paracellular permeability of intact monolayers with mouse embryonic stem cells

Abstract: Mouse embryonic stem (mES) cells were induced to form intact monolayers in cell culture inserts, using combinations of extracellular matrix (ECM) components and growth factors (GFs). Progressive formation of intact monolayers was monitored using transepithelial electrical resistance (TEER) and passage of paracellular permeability (PP) markers. The mES cells were initially inoculated on inactivated mouse embryonic fibroblasts (MEFs) plus leukemia inhibitory factor (LIF). At 75% confluence, cells were passaged i… Show more

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Cited by 26 publications
(12 citation statements)
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“…TEER measurements are used not only for epithelial barrier research but also for indications of the destruction of the extracellular matrix; for example, of neoplastic formations (Calabro et al 2008;Buchert et al 2012). To assess the effect of NPs on epithelial membrane integrity, we used the transepithelial resistance measurement of intact Caco2 and A549 epithelial cell monolayer.…”
Section: Resultsmentioning
confidence: 99%
“…TEER measurements are used not only for epithelial barrier research but also for indications of the destruction of the extracellular matrix; for example, of neoplastic formations (Calabro et al 2008;Buchert et al 2012). To assess the effect of NPs on epithelial membrane integrity, we used the transepithelial resistance measurement of intact Caco2 and A549 epithelial cell monolayer.…”
Section: Resultsmentioning
confidence: 99%
“…Based on TEER, sodium fluorescein permeation, and actin staining studies, DMSO, Cremophor EL, and PEG 400 (≤ 10%), as well as DMA and MPA (≤ 0.5%) appeared suitable for in vitro studies. Although TEER and paracelluar marker permeation are generally accepted indicators of tight junction permeability, these methods do not detect non-tight junction related epithelial membrane perturbation and as such are not good indicators of cell viability (Calabro et al, 2008). Therefore, more accurate indicators of cytotoxicity such as MTT and LDH were recommended as additional toxicity screening tools during in vitro studies (Sambruy et al, 2001).…”
Section: Discussionmentioning
confidence: 99%
“…Cultures were confluent at 7-days, but maximum resistance values (between 750–1000 Ω-cm 2 ) were achieved in intact monolayers after 15-days. Trans-membrane specific electrical resistance (TMER) was measured as previously described (Calabro et al ., 2008) using the Millicell-ERS® resistance system (Millipore) before and after 24-, 48- and 96-hr incubation with test chemicals. Blanks (inserts without cells containing media and chemical) were used to determine baseline values, and are expressed as percent TMER of untreated control groups.…”
Section: Methodsmentioning
confidence: 99%