Millicell ers mers00002

Manufactured by Merck Group

Sourced in United States, Germany

The Millicell-ERS is a laboratory instrument used for measuring electrical resistance and transepithelial electrical resistance (TEER) of cell monolayers. It provides a quantitative measurement of the integrity and tightness of cell culture models.

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Lab products found in correlation

Fitc dextran, by Merck Group (2 mentions) Transwell insert, by Corning (1 mentions) Bio one, by Greiner (1 mentions) Tnf α, by R&D Systems (1 mentions) Transwell insert, by BD (1 mentions)

Close spelling variants of this product

Millicell-ERS (153 citations) MERS00002 (17 citations) Millicells (10 citations)

11 protocols using millicell ers mers00002

Transepithelial Permeability Assay

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The upper and bottom chambers were filled with 250 μL and 800 μL of the medium, respectively, and cells were cultured as a monolayer on a cell culture insert (pore size 0.4 µm, high density, 353495; Corning Falcon). Values were recorded using Millicell-ERS (MERS00002; Merck Millipore), and the TEER value was obtained by subtracting the resistance of the corresponding naked filter from that of the cell monolayer filter.
For the permeability measurements, the culture medium was removed from the upper and bottom chambers, the upper chamber was replaced with 150 μL FITC-Dextran (1 mg/mL, MW 70000, 90718-1G; Sigma-Aldrich) diluted with medium, and the bottom chamber was replaced with 500 μL medium. After 20 min, 100 μL of the bottom chamber medium was sampled. The fluorescence of FITC-Dextran was measured using a multilabel counter (ARVO MX-1 1420; Perkin Elmer), and excitation/emission wavelengths were set at 485/535 nm. The fluorescence value of FITC-Dextran that passed through the cell culture insert without cells was shown to be 100%.

Takino J.I., Sato T., Kanetaka T., Okihara K., Nagamine K., Takeuchi M, & Hori T. (2021). RasGRP2 inhibits glyceraldehyde-derived toxic advanced glycation end-products from inducing permeability in vascular endothelial cells. Scientific Reports, 11, 2959.

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Endothelial Barrier Integrity Assay

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Endothelial barrier integrity was determined using by the electrical resistance system (Millicell‐ERS, MERS00002; Merck Millipore, Germany). Cells were treated as we described previously, and then, electric resistance of monolayer HUVECs was measured every 4 hours; Transendothelial electric resistance (TEER) was calculated based on the manufacturer's recommendations. Then, TEER was graphed at indicated time‐points during an experiment at 24 hours. A higher TEER represented a higher barrier integrity.

Zheng X., Zhang W, & Wang Z. (2019). Simvastatin preparations promote PDGF‐BB secretion to repair LPS‐induced endothelial injury through the PDGFRβ/PI3K/Akt/IQGAP1 signalling pathway. Journal of Cellular and Molecular Medicine, 23(12), 8314-8327.

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Measuring Epithelial Barrier Integrity

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Epithelial barrier integrity was determined by measurement of electrical resistance (Millicell-ERS, MERS00002; Merck Millipore, Darmstadt, Germany). Cells were seeded in Transwell inserts (pore size 0.4 μm, #3470; Costar) as previously described; the electric resistance of A549 cell monolayers was measured at 2-h intervals. Transepithelial electric resistance (TEER) was calculated in accordance with the manufacturer’s recommendations; a higher TEER was indicative of greater barrier integrity.

Zhang W., Zhou H., Jiang Y., He J., Yao Y., Wang J., Liu X., Leptihn S., Hua X, & Yu Y. (2022). Acinetobacter baumannii Outer Membrane Protein A Induces Pulmonary Epithelial Barrier Dysfunction and Bacterial Translocation Through The TLR2/IQGAP1 Axis. Frontiers in Immunology, 13, 927955.

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Transwell Assay for Podocyte Barrier Assessment

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Cellular barrier properties were assessed using assessment of TER across confluent podocytes with Millicell-ERS (MERS00002, Millipore, Darmstadt, Germany) [23 (link)]. A 24-well transwell system (Greiner Bio-One, 0.4-mm pore size, 6.5-mm diameter, transparent, Costar) was inserted into the 24-well plate. Podocytes were plated in transwell chambers. The volumes of culture medium were 100 and 600 μL in the upper and lower compartments, respectively, in which cells were allowed to grow for 2 days (considered day 0). The resistance (R sample) was measured; the value for the blank well represented the blank resistance (R blank). The following calculation was performed: epithelial monolayer resistance = (R sample-R blank) * effective membrane area (the area of a 24-well transwell chamber is 0.336 cm²).

Li L., Feng Y., Zhang J., Zhang Q., Ren J., Sun C., Li S., Lei X., Luo G., Hu J, & Huang Y. (2022). Microtubule associated protein 4 phosphorylation-induced epithelial-to-mesenchymal transition of podocyte leads to proteinuria in diabetic nephropathy. Cell Communication and Signaling : CCS, 20, 115.

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Transwell System for Endothelial Barrier Evaluation

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The cellular barrier properties were assessed by measuring the TER across confluent HPMECs using Millicell-ERS (MERS00002, Millipore, USA)38 (link). The 24-well Transwell system was inserted into the 24-well plate. HPMECs were plated in Transwell chambers; the volumes of culture medium in the upper and lower compartments were 100 and 600 μl, respectively, and the cells were grown for 4 days and replaced with serum-free medium for 1 hr before treatment. The electrode tips were inserted into the upper compartment and the lower compartment, respectively, and the resistance (R sample) was measured; the value for the blank well represented the blank resistance (R blank). The following calculation was performed: endothelial monolayer resistance = (R sample-R blank)*effective membrane area (the area of a 24-well Transwell chamber is 0.336 cm²).

Li L., Hu J., He T., Zhang Q., Yang X., Lan X., Zhang D., Mei H., Chen B, & Huang Y. (2015). P38/MAPK contributes to endothelial barrier dysfunction via MAP4 phosphorylation-dependent microtubule disassembly in inflammation-induced acute lung injury. Scientific Reports, 5, 8895.

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HUVEC Transwell Permeability Assay

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HUVECs were seeded in the top chambers of 24-well Transwell plates (8.0-µm pore size; Cat. No. 35-3097; Becton-Dickinson, Franklin Lakes, NJ, United States) at a density of 1 × 10⁴ cells/well. Volumes of culture media in the top and lower chambers were 300 and 700 μL, respectively. TEER was measured using a Millicell-ERS (MERS00002; Millipore, Burlington, MA, United States), as previously described (Li et al., 2015 (link)). Values for blank wells and cell wells represented the blank resistance (R blank) and sample resistance (R Sample), respectively. The following calculation was performed: TEER = (R Sample)–(R blank). The effect of IH on TEER was determined in HUVECs after 9, 18, 36, 72, and 108 cycles of IH exposure. The effect of drugs on permeability was determined in HUVECs pretreated with salidroside (10 µM or 100 µM), H-89 (10 μM), Y27632 (10 µM), or Fasudil (10 µM) for 2 h before exposure to either normoxia or 72 cycles of IH.

Li L., Yang Y., Zhang H., Du Y., Jiao X., Yu H., Wang Y., Lv Q., Li F., Sun Q, & Qin Y. (2021). Salidroside Ameliorated Intermittent Hypoxia-Aggravated Endothelial Barrier Disruption and Atherosclerosis via the cAMP/PKA/RhoA Signaling Pathway. Frontiers in Pharmacology, 12, 723922.

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Measuring Endothelial Barrier Integrity

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The cellular barrier properties were assessed by measuring the resistance values across confluent HPMECs using Millicell‐ERS (MERS00002, Millipore; Monaghan‐Benson & Wittchen, 2011 (link)). The 12‐well Transwell system was inserted into the 12‐well plate. HPMECs were plated in Transwell chambers. The electrode tips were inserted into the upper and lower compartments, and the resistance (R sample) was measured. The value for the blank well represented the blank resistance (R blank). The transendothelial electrical resistance (TEER) value was calculated based on the resistance per unit area as (R sample − R blank) × 1.33 cm (the area of a 12‐well Transwell chamber is 1.33 cm²; Tominaga et al., 2015 (link)).

Kuang X., Wang Y., Liu S., Chang L., Yin Y., Li Z., Liu Y., Li W., Hou Y., Wang H., Liang J, & Jia Z. (2022). Tongxinluo enhances the effect of atorvastatin on the treatment of atherosclerosis with chronic obstructive pulmonary disease by maintaining the pulmonary microvascular barrier. Food Science & Nutrition, 11(1), 390-407.

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In Vitro BBB Co-Culture Model

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The in vitro BBB co-culture model was constructed in the transwell chamber following methods previously reported [28 (link)]. In brief, hBMECs were pre-seeded in the up chamber of transwell inserts (6.5 mm diameter inserts, 3.0 μm pore size) at 8 × 10³ cells per well, and U251 astrocytes were synchronously seeded in the 24-wells plate. After 12 h of growth, the transwell inserts with hBMECs were transferred to the 24-wells plate containing U251, and the co-culture medium was changed into the hBMECs complete medium. During co-cultivation, the TEER values were detected with Millicell-ERS (MERS00002) instrument (Millipore, Burlington, MA, USA).

Fu J., Li L., Huo D., Zhi S., Yang R., Yang B., Xu B., Zhang T., Dai M., Tan C., Chen H, & Wang X. (2021). Astrocyte-Derived TGFβ1 Facilitates Blood–Brain Barrier Function via Non-Canonical Hedgehog Signaling in Brain Microvascular Endothelial Cells. Brain Sciences, 11(1), 77.

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Endothelial Barrier Permeability Dynamics

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Changes of the transendothelial electrical resistance (TEER) of endothelial cells were measured with the Millicell-ERS (MERS00002, Millipore, Bedford, MA, USA). An in vitro transendothelial permeability assay was performed quantifying transendothelial passage of fluorescein isothiocyanate-dextran (FITC-dextran, 40-kDa; Sigma). HPMECs were seeded at a density of 1.5 × 10⁵cells/well on 24-well Transwell plates (Greiner Bio-One, pore size: 0.4 mm; diameter: 6.5 mm, Costar, The Netherlands), and were stimulated with 10 μg/mL of LPS or 10 ng/mL of TNF-α (R&D Systems, Abingdon, UK). After stimulation for 1, 6, 12, and 24 h, the TEER across the HPMECs was measured. To evaluate the effect of UFH on HPMEC hyperpermeability induced by LPS or TNF-α, the cells were stimulated by 10 μg/mL of LPS or 10 ng/mL of TNF-α for 6 h after pre-treatment with vehicle or UFH (10 U/mL). The measurements were performed as previously described.^{[24 (link)]}

Mu S.T., Tang J., Ma J.Q., Zhong Y., Liu H.Z., Ma X.C, & Zheng Z. (2020). Unfractionated heparin attenuates endothelial barrier dysfunction via the phosphatidylinositol-3 kinase/serine/threonine kinase/nuclear factor kappa-B pathway. Chinese Medical Journal, 133(15), 1815-1823.

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Determining Airway Cell Polarity by TEER

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During culture of WD cells in differentiation medium, the polarity of cells was determined by TEER measurement. The apical and basolateral chambers of inserts were filled with fresh differentiation medium following 21 days of ALI culture, and equilibrated at 37 °C, 5% CO2 for 10 min. TEER values were determined using two Millicell-ERS (MERS00002, Millipore, Burlington, MA, USA) electrodes submerged into the insert medium. WD cells with TEER values > 1000 Ω cm² were considered well-differentiated and met the requirement for subsequent studies of the model (Hiemstra et al., 2018 (link); Huang et al., 2012 ; Min et al., 2016 (link)).

Liu W.K., Xu D., Xu Y., Qiu S.Y., Zhang L., Wu H.K, & Zhou R. (2020). Protein profile of well-differentiated versus un-differentiated human bronchial/tracheal epithelial cells. Heliyon, 6(6), e04243.

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