Established human renal cell lines: Phenotypic characteristics define suitability for use in in vitro models for predictive toxicology

In renal medullas during antidiuresis, the extracellular fluid is hyperosmotic because of high concentrations of NaCl and urea. Under those conditions, the cells contain high concentrations of organic osmolytes, namely sorbitol, myo-inositol, glycerophosphorylcholine (GPC), and betaine to balance the extracellular hyperosmolality. These organic osmolytes increase cell osmolality without perturbing the intracellular milieu in ways that would degrade the function of cellular macromolecules. The present study surveyed a number of cell lines for the ability to survive in media with high concentrations of NaCl and/or urea and for the accumulation of organic osmolytes. Of the renal cell lines tested, MDCK, GRB-MAL1, and A6 cells proliferated in hyperosmotic media, but medullary interstitial cells LLC-PK1 and LLC-PK3 did not proliferate, nor did nonrenal HTC-BH cells, MDCK, LLC-PK1, and LLC-PK3 cells contained higher concentrations of myo-inositol, GPC, and betaine when cultured in media containing high NaCl (with or without high urea) and much lower or undetectable levels of these osmolytes when grown in isosmotic media. Sorbitol, and to a lesser extent myo-inositol, were elevated in GRB-MAL1 cells in media hyperosmotic with NaCl but not in isosmotic media. There was less accumulation of organic osmolytes when only urea was added to increase osmolality. Thus the same osmolytes were accumulated by one or another cell line in vitro as were previously found in renal medullas. These cell lines provide models for studying osmolyte accumulation.

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1H NMR spectroscopic studies on the characterization of renal cell lines and identification of novel potential markers of in vitro nephrotoxicity

Biomarkers ◽

10.3109/13547509609079345 ◽

1996 ◽

Vol 1 (1) ◽

pp. 35-43 ◽

Cited By ~ 3

Author(s):

Maria L. Anthony ◽

Peter C. R. McDowell ◽

Tim J. B. Gray ◽

Melanie Blackmore ◽

Jeremy K. Nicholson

Keyword(s):

Cell Lines ◽

1H Nmr ◽

Renal Cell ◽

Spectroscopic Studies ◽

Renal Cell Lines

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RENAL CELL LINES FOR STUDY OF NEPHROTOXICITY IN VITRO

Military Medical Science Letters ◽

10.31482/mmsl.2016.013 ◽

2016 ◽

Vol 85 (2) ◽

pp. 69-74 ◽

Cited By ~ 1

Author(s):

Martina Vrbová ◽

Eva Dastychová ◽

Tomáš Roušar

Keyword(s):

Cell Lines ◽

Renal Cell ◽

Renal Cell Lines

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Defining candidate mRNA and protein EV biomarkers to discriminate ccRCC and pRCC from non-malignant renal cells in vitro

Medical Oncology ◽

10.1007/s12032-021-01554-2 ◽

2021 ◽

Vol 38 (9) ◽

Author(s):

Richard C. Zieren ◽

Liang Dong ◽

David J. Clark ◽

Morgan D. Kuczler ◽

Kengo Horie ◽

...

Keyword(s):

Cell Lines ◽

Renal Cell ◽

Messenger Rna ◽

Size Exclusion ◽

Tandem Mass Tag ◽

Exclusion Chromatography ◽

Promising Source ◽

Renal Cell Lines ◽

Pathological Group

AbstractRenal cell carcinoma (RCC) accounts for over 400,000 new cases and 175,000 deaths annually. Diagnostic RCC biomarkers may prevent overtreatment in patients with early disease. Extracellular vesicles (EVs) are a promising source of RCC biomarkers because EVs carry proteins and messenger RNA (mRNA) among other biomolecules. We aimed to identify biomarkers and assess biological functions of EV cargo from clear cell RCC (ccRCC), papillary RCC (pRCC), and benign kidney cell lines. EVs were enriched from conditioned cell media by size exclusion chromatography. The EV proteome was assessed using Tandem Mass Tag mass spectrometry (TMT-MS) and NanoString nCounter technology was used to profile 770 cancer-related mRNA present in EVs. The heterogeneity of protein and mRNA abundance and identification highlighted the heterogeneity of EV cargo, even between cell lines of a similar pathological group (e.g., ccRCC or pRCC). Overall, 1726 proteins were quantified across all EV samples, including 181 proteins that were detected in all samples. In the targeted profiling of mRNA by NanoString, 461 mRNAs were detected in EVs from at least one cell line, including 159 that were present in EVs from all cell lines. In addition to a shared EV cargo signature, pRCC, ccRCC, and/or benign renal cell lines also showed unique signatures. Using this multi-omics approach, we identified 34 protein candidate pRCC EV biomarkers and 20 protein and 8 mRNA candidate ccRCC EV biomarkers for clinical validation.

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Endogenous H2S production deficiencies lead to impaired renal erythropoietin production

Canadian Urological Association Journal ◽

10.5489/cuaj.5658 ◽

2018 ◽

Vol 13 (7) ◽

Cited By ~ 2

Author(s):

Jennifer Leigh ◽

Smriti Juriasingani ◽

Masoud Akbari ◽

Peng Shao ◽

Manujendra N. Saha ◽

...

Keyword(s):

Cell Lines ◽

Renal Cell ◽

Oxygen Sensor ◽

Whole Body ◽

Clinical Samples ◽

Biosynthetic Enzyme ◽

Wild Type ◽

Renal Cell Lines

Introduction: Patients suffering from chronic kidney disease (CKD) experience a number of associated comorbidities, including anemia. Relative deficiency in renal erythropoietin (EPO) production is thought to be a primary cause of anemia. Interestingly, CKD patients display low levels of hydrogen sulfide (H2S), an endogenously derived renal oxygen sensor. Previous in vitro experiments have revealed that H2S-deficient renal cell lines produce less EPO than wild-type renal cell lines during hypoxia.Methods: We postulated that H2S might be a primary mediator of EPO synthesis during hypoxia, which was tested using an in vivo murine model of whole-body hypoxia and in clinical samples obtained from CKD patients.Results: Following a 72-hour period of hypoxia (11% O2), partial H2S knockout mice (lacking the H2S biosynthetic enzyme cystathionine γ-lyase [CSE]) displayed lower levels of hemoglobin, EPO and cystathionine-β-synthase (CBS) (another H2S biosynthetic enzyme) compared to wild-type mice, all of which was rescued by exogenous H2S supplementation. We also found that anemic CKD patients requiring exogenous EPO exhibited lower urinary thiosulfate levels compared to non-anemic CKD patients of similar CKD classification.Conclusions: Together, our results confirm an interplay between the actions of H2S during hypoxia and EPO production.

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Malignant hematopoietic cell lines: In vitro models for the study of primary mediastinal B-cell lymphomas

Leukemia Research ◽

10.1016/j.leukres.2014.11.002 ◽

2015 ◽

Vol 39 (1) ◽

pp. 18-29 ◽

Cited By ~ 8

Author(s):

Hans G. Drexler ◽

Stefan Ehrentraut ◽

Stefan Nagel ◽

Sonja Eberth ◽

Roderick A.F. MacLeod

Keyword(s):

B Cell ◽

Cell Lines ◽

In Vitro Models ◽

Hematopoietic Cell ◽

B Cell Lymphomas ◽

Hematopoietic Cell Lines

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Hepatoma Cell Cultures as In Vitro Models for the Hepatotoxicity of Xenobiotics

Alternatives to Laboratory Animals ◽

10.1177/026119298801600108 ◽

1988 ◽

Vol 16 (1) ◽

pp. 32-37

Author(s):

Margherita Ferro ◽

Anna Maria Bassi ◽

Giorgio Nanni

Keyword(s):

Cell Lines ◽

Cell Cultures ◽

Membrane Lipids ◽

Hepatoma Cell ◽

Positive Control ◽

In Vitro Models ◽

Low Concentrations ◽

Formation Efficiency ◽

Dose Dependent

Two hepatoma cell cultures were examined as in vitro models to be used in genotoxicity and cytotoxicity tests without the addition of bioactivating enzymes. The MH1C1, and HTC hepatoma lines were used in this study to establish their sensitivity to a number of xenobiotics, namely, cyclophosphamide (CP), the classical positive control in bioactivation tests; benzaldehyde (BA), a short-chain aldehyde; and 4-hydroxynonenal (HNE), a major toxic end-product of the peroxidative degradation of cell membrane lipids. As a first approach, we compared the following cytotoxicity tests: release of lactate dehydrogenase (LDH), and colony formation efficiency (CF). Colony-forming cells were exposed to the drugs according to different procedures, before or after the anchorage phase. The leakage of LDH into the medium following exposure of both cell lines to HNE, CP and BA for up to 24 hours was found not to be a good index of cytotoxicity. A better indicator of cytotoxicity was CF, as evaluated by exposure of the cells 24 hours after seeding. The effects were detectable at very low concentrations, corresponding to 10, 90 and 100μM for HNE, CP and BA, respectively. The impairment of CF efficiency was dose-dependent and time-dependent, and several differences between the two cell lines were observed.

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Upregulation of FBXW7 Suppresses Renal Cancer Metastasis and Epithelial Mesenchymal Transition

Disease Markers ◽

10.1155/2017/8276939 ◽

2017 ◽

Vol 2017 ◽

pp. 1-7 ◽

Cited By ~ 6

Author(s):

Yangke Cai ◽

Meng Zhang ◽

Xiaofu Qiu ◽

Bingwei Wang ◽

Yu Fu ◽

...

Keyword(s):

Cell Lines ◽

Renal Cell ◽

Cancer Metastasis ◽

Epithelial Mesenchymal Transition ◽

Migration And Invasion ◽

Coding Region ◽

Mesenchymal Transition ◽

Renal Cell Line ◽

Emt Markers

Background and Objective. FBXW7, known as a general tumor suppressor, is commonly lowly expressed in metastatic malignancies. We aim to investigate the potential influence of FBXW7 overexpression on renal cell carcinoma (RCC) metastasis. Methods. We employed quantitative real-time PCR (qRT-PCR) and Western blotting (WB) to quantify the FBXW7 expression in RCC cell lines. Upregulation of FBXW7 was performed in vitro on RCC cells using the lentivirus covering coding region FBXW7 cDNA sequence, and functional tests were performed to verify FBXW7 overexpression on migration and invasion of RCC cells. Moreover, WB was employed to determine the expressions of MMP-2, MMP-9, and MMP-13, as well as EMT markers in the transfected RCC cells. Results. FBXW7 was significantly downregulated in RCC cell lines, dominated by 786-O and ACHN, when compared to normal renal cell line HK-2. Moreover, upregulation of FBXW7 in 786-O and ACHN cell lines significantly inhibited cell migration and invasion, as well as EMT. Present study also showed that FBXW7 was involved in the migration and invasion of RCC cells via regulating the expressions of MMP-2, MMP-9, and MMP-13. Conclusion. Our findings demonstrate that upregulation of FBXW7 inhibits RCC metastasis and EMT. FBXW7 is a potential therapeutic target for RCC patients.

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