Transepithelial Resistance and Inulin Permeability as Endpoints in <i>In Vitro</i> Nephrotoxicity Testing

Duff, Tracey; Carter, Simon; Feldman, Gemma; McEwan, Gordon T.A.; Pfaller, Walter; Rhodes, Pauline; Ryan, Michael P.; Hawksworth, Gabrielle

doi:10.1177/026119290203002s08

Cited by 24 publications

(12 citation statements)

References 6 publications

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“…Phosphate‐buffered saline (PBS, 1 mL) was placed inside the Endohm chamber, the culture inserted atop and 150 µL of PBS was added apically into the insert and the measurement taken. Prior to testing the cultures TEER an empty culture insert (containing 150 µL PBS) was used as a “blank” and subtracted from each subsequent sample reading 45. TEER measurements were taken on day 1 and every third day up to day 42 for all 3 donors A, B and C ( n = 3) and are presented as resistance values (in Ω · cm 2 ).…”

Section: Methodsmentioning

confidence: 99%

Tissue‐Specific Stem Cell Differentiation in an in vitro Airway Model

Prytherch

Job

Marshall

et al. 2011

Macromolecular Bioscience

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The respiratory tract is the primary site of exposure to airborne compounds, with the bronchial epithelium providing one of the first lines of defence. A growing need exists for an accurate in vitro model of the bronchial epithelium. Here, normal human bronchial epithelial (NHBE) cells cultured at an air/liquid interface create a fully differentiated, in-vivo-like model of the human bronchial epithelium. Developmental characterisation includes (i) trans-epithelial electrical resistance, (ii) morphology and (iii) bronchial cell specific stains/markers. It is concluded that the basal/progenitor cells create a pseudo-stratified, mucociliary NHBE model containing basal, serous, Clara, goblet and ciliated cells, reflective of the normal human bronchial epithelium (days 24-33 ALI culture).

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Section: Methodsmentioning

confidence: 99%

Tissue‐Specific Stem Cell Differentiation in an in vitro Airway Model

Prytherch

Job

Marshall

et al. 2011

Macromolecular Bioscience

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“…However, the latter was carried out primarily with a focus on transcriptomic, metabolomic, and proteomic approaches and not with a focus on functional characteristics. While functional readouts are commonly applied for the in vitro detection of neuro- (Hausherr et al 2014) or cardiotoxicity (Sirenko et al 2017), such approaches have so far been rarely applied for the detection of nephrotoxicity and, when applied, were basically limited to measurements of transepithelial electrical resistance (Duff et al 2002). Thus, in the present study, we investigated whether the combination of functional measurements, e.g., water reabsorption and cation secretion with classical cytotoxicity endpoints, while using a limited set of nephrotoxic and non-nephrotoxic reference substances, could provide for improved sensitivity in the detection of drug-induced nephrotoxicity.…”

Section: Introductionmentioning

confidence: 99%

Functional transepithelial transport measurements to detect nephrotoxicity in vitro using the RPTEC/TERT1 cell line

et al. 2019

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The kidney is a frequent target for organ-specific toxicity as a result of its primary function in controlling body fluids, for example, via resorption of amino acids, peptides, nutrients, ions, xenobiotics and water from the primary urine as well as excretion of metabolic waste products and hydrophilic and amphiphilic xenobiotics. Compounds exhibiting dose-limiting nephrotoxicity include drugs from highly diverse classes and chemical structures, e.g., antibiotics (gentamicin), chemotherapeutics (cisplatin), immunosuppressants (cyclosporine A and tacrolimus) or bisphosphonates (zoledronate). All of these compounds elicit nephrotoxicity primarily by injuring renal proximal tubule epithelial cells (RPTECs). However, prediction of a compound's nephrotoxic potential in humans to support early unmasking of risk-bearing drug candidates remains an unmet challenge, mainly due to the complex kidney anatomy as well as pronounced inter-and intraspecies differences and lack of relevant and validated human in vitro models. Accordingly, we used the recently established human RPTEC/TERT1 cell line to carry out toxicity studies with a focus on impairment of functional characteristics, i.e., transepithelial electrical resistance (TEER), vectorial transport of water, cations, and anions. Results were compared to real-time cytotoxicity assessments using cellular impedance (xCELLigence assay) and the routine cell viability readout (MTT). As expected, most toxins caused exposure time-and concentration-dependent cytotoxicity. However, for some compounds (cyclosporine A and tacrolimus), transport processes were strongly impaired in absence of a concomitant decrease in cell viability. In conclusion, these data demonstrate that functional parameters are important, highly sensitive and meaningful additional readouts for nephrotoxicity assessment in human renal proximal tubule epithelial cells.

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“…However, the use of different technical approaches to measure TER sometimes results in inconsistent reports for TER readings within the same cell lines. For example, when using chopstick electrodes the TER values obtained were higher in A6 renal cells [11], MDCK strain 1 and LLCK1 porcine kidney cells [12] than those obtained using Endohm chambers. These higher values may be attributable to incorrect positioning of the chopstick electrodes, resulting in the subjection of cells to a non-uniform electrical field [11].…”

Section: Discussionmentioning

confidence: 99%

Comparison of transepithelial resistance measurement techniques: Chopsticks vs. Endohm

2017

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BackgroundTER measurements across confluent cellular monolayers provide a useful indication of TJ strength between epithelial and endothelial cells in culture. Having a reliable and accurate method of measuring cell-to-cell adhesion is critical to studies in pathophysiology and cancer metastasis. However, the use of different technical approaches to measure TER has reportedly yielded inconsistent measurements within the same cell lines.MethodsIn the current study, we compared the peak TER values for the MDCK (canine kidney) and MCF-7 (human breast cancer) epithelial cell lines using two common approaches (Chopstick and Endohm) and two types of polymer inserts (PC and PET).ResultsBoth cell lines demonstrated a statistically significant difference in the peak TERs obtained using the two different approaches. Further, the MDCK (but not the MCF-7) cells demonstrated a statistically significant difference between the peak TERs when using the same approach but different inserts.ConclusionOur study indicates the importance of using a single approach when seeking to measure and compare the TER values of cultured cell lines.

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Transepithelial Resistance and Inulin Permeability as Endpoints in In Vitro Nephrotoxicity Testing

Cited by 24 publications

References 6 publications

Tissue‐Specific Stem Cell Differentiation in an in vitro Airway Model

Tissue‐Specific Stem Cell Differentiation in an in vitro Airway Model

Functional transepithelial transport measurements to detect nephrotoxicity in vitro using the RPTEC/TERT1 cell line

Comparison of transepithelial resistance measurement techniques: Chopsticks vs. Endohm

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