Eksempler på brug af Teer på Engelsk og deres oversættelser til Dansk
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I don't know how that's gonna happen, Teer.
TEER, leakage assays, and tight junction staining.
Transepithelial electrical resistance(TEER) was measured to assess monolayer integrity.
TEER values in the absence of NCs are shown as controls the dashed interval marks S.E.M of the mean value.
Validation of the GI Epithelial Permeability Barrier Using Transepithelial Electrical Resistance(TEER) and Immunostaining of Tight Junctions.
Use wells with low TEER as a negative control for barrier formation.
Add 125I-antibody NCs,(e.g.125I-anti-ICAM NCs; 56 nCi/ml) to the upper chamber above confluent Caco-2 monolayers with TEER≥350 Ω×cm2(see Discussion) over background 16-21 days post-seeding.
Measure TEER(Section 2.3) before and after incubation to assess the effects of NCs on the permeability barrier.
Formation of a permeability barrier is confirmed by measuring transepithelial electrical resistance(TEER), transepithelial transport of a control substance, and immunostaining of tight junctions.
Plateau TEER values and time necessary to achieve barrier integrity may vary according to cell passage number.
Calculate resistivity(resistance normalized to area; Ω×cm2) of samples by subtracting background resistance(TEER of transwell inserts without cells) and multiplying by membrane surface area.
Measure TEER(Section 2.3) before, during, and after incubation, then collect all fractions for total 125I and free 125I measurements as indicated in Section 3.
Culturing of GI epithelial cell monolayers in transwell inserts enabled measurement of TEER and fluorescence immunostaining of tight junctions to confirm formation of a cell permeability barrier.
Measure TEER at various time points throughout the incubation to identify TEER value decay caused by H2O2-induced opening of the cell junctions.
As a platform to study transepithelial transport, Caco-2 cell monolayers are cultured on transwell inserts until a tight monolayer is formed 16-21 days post-seeding, TEER≥ 350 Ω×cm2 over background.
However, incubation with anti-ICAM NCs did not alter TEER or 125I-albumin paracellular leakage over a period of 48 hr, similar to control IgG NCs that are not transported.
Once a transwell model is established, including the appropriate selection of cell type, media composition, and permeable filter, the status of the cell monolayer can be assessed during the culturing andexperimental stages using TEER.
Measure TEER(Section 2.3) before, during, and after incubation of cells with inhibitors and radiolabeled materials, as an additional control for the effect of the inhibitors on the monolayer permeability.
To verify the presence of tight junctions in cell monolayers with high TEER(equal to or above the threshold value for barrier formation), fix cells by incubation with cold 2% paraformaldehyde for 15 min.
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Finally, mechanistic studies were applied using endocytic inhibitors, TEER, and an albumin paracellular leakage assay to assess whether transport occurs through a vesicular route versus a paracellular route.
As validation of our cell model to study transepithelial transport of targeted NCs, Figure 2 shows that Caco-2 cell monolayers plated at a density of 1.5× 105 cells/cm2 reached confluence~Day 12 andmaintained monolayer integrity up to Day 18, indicated by TEER Figure 2A.
Repeat measurements every 1-2 days until TEER values rise to a maximum and plateau, indicating formation of a permeability barrier, which typically takes 2-3 weeks from the moment of cell platting.
In terms of assessing the mechanism of transport,the same methods described for validating monolayer integrity(TEER, leakage assays, and tight junction staining) can be used to evaluate the paracellular route.
To verify the presence of tight junctions in cell monolayers with high TEER(equal to or above the threshold value for barrier formation), fix cells by incubation with cold 2% paraformaldehyde for 15 min. Then, wash cells with PBS and incubate them for 30 min at room temperature with 1 μg/ml anti-occludin.
This was validated by the presence of occludin-positive tight junctions(Figure 2B) in monolayers with high TEER(390 Ω×cm2, Day 14), compared to poor tight junction labeling at low TEER 17 Ω×cm2, Day 5.
Figure 1. Schematic of transepithelial transport of targeted nanocarriers across a gastrointestinal epithelial model.(A) As a platform to study transepithelial transport, Caco-2 cell monolayers are cultured on transwell inserts until a tight monolayer is formed 16-21 days post-seeding, TEER≥ 350 Ω×cm2 over background.
The status of the permeability barrier is validated by measuring TEER, apical-to-basolateral transport of a control substance, albumin, and fluorescence microscopy visualization of an element of the tight junctions, occludin protein.
Supplementary to this is the use of controls for non-specific staining,using fluorescent secondary antibodies only, absence of tight junctions(here, we used cells with a monolayer showing low TEER), or disruption of tight junctions using the junction-modulating agents listed above.
Due to the various biological andenvironmental factors affecting TEER, a minimum baseline value indicating barrier formation must be established for each system by regularly monitoring TEER during the culture period and after experimental manipulations.