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PODO/TERT256 : A promising human immortalized podocyte cell line and its potential use for in vitro research at different oxygen levels

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2024

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Chemico-Biological Interactions. Elsevier. 2024, 387, 110813. ISSN 0009-2797. eISSN 1872-7786. Available under: doi: 10.1016/j.cbi.2023.110813

Zusammenfassung

Podocytes are of key interest for the prediction of nephrotoxicity as they are especially sensitive to toxic insults due to their central role in the glomerular filtration apparatus. However, currently, prediction of nephrotoxicity in humans remains insufficiently reliable, thus highlighting the need for advanced in vitro model systems using human cells with improved prediction capacity. Recent approaches for refining in vitro model systems focus on closely replicating physiological conditions as observed under the in vivo situation typical of the respective nephron section of interest. PODO/TERT256, a human immortalized podocyte cell line, were employed in a semi-static transwell system to evaluate its potential use as a human podocyte in vitro system for modelling potential human glomerular toxicity. Furthermore, the impact of routinely employed excessive oxygen tension (21 % - AtmOx), when compared to the physiological oxygen tensions (10 % - PhysOx) observed in vivo, was analyzed. Generally, cultured PODO/TERT256 formed a stable, contact-inhibited monolayer with typical podocyte morphology (large cell body, apical microvilli, finger-like cytoplasmic projections (reminiscent of foot processes), and interdigitating cell-cell junctions) and developed a size-selective filtration barrier. PhysOx, however, induced a more pronounced in vivo like phenotype, comprised of significantly larger cell bodies, significantly enhanced filtration barrier size-selectivity, and a remarkable re-localization of nephrin to the cell membrane, thus suggesting an improved in vitro replication of in vivo characteristics. Preliminary toxicity characterization with the known glomerulotoxin doxorubicin (DOX) suggested an increasing change in filtration permeability, already at the lowest DOX concentrations tested (0.01 μM) under PhysOx, whereas obvious changes under AtmOx were observed as of 0.16 μM and higher with a near all or nothing effect. The latter findings suggested that PODO/TERT256 could serve as an in vitro human podocyte model for studying glomerulotoxicity, whereby culturing at PhyOx tension appeared critical for an improved in vivo-like phenotype and functionality. Moreover, PODO/TERT256 could be incorporated into advanced human glomerulus systems in vitro, recapitulating microfluidic conditions and multiple cell types (endothelial and mesenchymal cells) that can even better predict human glomerular toxicity.

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570 Biowissenschaften, Biologie

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ISO 690SCHLICHENMAIER, Nadja, Alexander ZIELINSKI, Sascha BENEKE, Daniel R. DIETRICH, 2024. PODO/TERT256 : A promising human immortalized podocyte cell line and its potential use for in vitro research at different oxygen levels. In: Chemico-Biological Interactions. Elsevier. 2024, 387, 110813. ISSN 0009-2797. eISSN 1872-7786. Available under: doi: 10.1016/j.cbi.2023.110813
BibTex
@article{Schlichenmaier2024PODOT-68956,
  year={2024},
  doi={10.1016/j.cbi.2023.110813},
  title={PODO/TERT256 : A promising human immortalized podocyte cell line and its potential use for in vitro research at different oxygen levels},
  volume={387},
  issn={0009-2797},
  journal={Chemico-Biological Interactions},
  author={Schlichenmaier, Nadja and Zielinski, Alexander and Beneke, Sascha and Dietrich, Daniel R.},
  note={Article Number: 110813}
}
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