![]() ![]() This assay had moderate correlation with the TER, another measure of permeability which measures resistance to ion flow across the cell layer. By fluorescence-based cell culture assay, hTERT immortalized corneal epithelium had a tighter barrier than hTERT immortalized conjunctival epithelium, such as in the in vivo situation. ![]() The TER was measured using endohm electrodes and an EVOM voltohmmeter (World Precision Instruments, LTD., Hertfordshire, UK).Īfter 12 days in culture, the permeability of FITC-Dextran was 0.0188µl/cm 3/min for corneal cells and 0.0398µl/cm 3/min for conjunctival cells.FITC-Dextran permeability assay and TER had moderate negative correlation (correlation coefficient: -0.44).Ĭorneal and conjunctival epithelium forms a barrier that isolates the eye from the outside environment and regulates passive movement through the paracellular pathway. ![]() Up to a 1-year Warranty & Lifetime Support. A standard curve was created based upon fixed concentrations of fluorescence. Refurbished WPI World Precision Instruments EVOM 2 Epithelial Voltohmmeter, tested & serviced prior to shipping. All values are expressed as percentages of the basal level for the controls. TEER values were obtained by subtracting the contribution of the filter and bathing solution. Bioz Stars score: 86/100, based on 1 PubMed. TEER was determined with the use of an EVOM voltohmmeter connected to a 12-mm Endohm unit (World Precision Instruments). The fluorescence of each sample was determined in a fluorescence spectrofluorometer at an excitation wavelength of 485 nm and an emission wavelength of 535 nm. Evom Epithelial Tissue Voltohmmeter, supplied by World Precision Instruments, used in various techniques. Aliquots of 30µl were taken from cell cultures at 30, 60, and 90 min. Bioz Stars score: 86/100, based on 1 PubMed citations. Four mg/ml fluorescein isothiocyanate (FITC) Dextran 4000-conjugate (MW4000) was added to the apical medium chamber. Evom Epithelial Voltammeter, supplied by World Precision Instruments, used in various techniques. Permeability assays were performed after 12 days in culture, when the TER (transepithelial electrical resistance) had reached a plateau. The morphology of the cultures was assessed by labeling actin filaments or tight junction-related proteins by indirect immunofluorescence. To create a reproducible and accurate fluorescence-based cell culture assay and evaluate the barrier function in hTERT immortalized conjunctival and corneal epithelium.Ĭonjunctival and corneal epithelial cells immortalized by preventing telomere shortening by transduction with hTERT were cultured on 12-mm Transwell filters at a density of 4 x 10 5 cells/cm 2. ![]()
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