Dokument: Influence of Nephrotoxins on the Differentiation Process of Human Induced Pluripotent Stem Cells (hiPSC) to Proximal Tubular Epithelial-Like Cells (PTELC)
| Titel: | Influence of Nephrotoxins on the Differentiation Process of Human Induced Pluripotent Stem Cells (hiPSC) to Proximal Tubular Epithelial-Like Cells (PTELC) | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=71617 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20251208-094806-9 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Kouidrat, Nazih Mohamed Zakari [Autor] | |||||||
| Dateien: |
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| Beitragende: | Schupp, Nicole [Gutachter] Adjaye, James [Gutachter] | |||||||
| Dewey Dezimal-Klassifikation: | 600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit | |||||||
| Beschreibung: | Proximal tubule epithelial cells (PTEC) are the most metabolically active renal segment in solute reabsorption and drug metabolism and therefore the most vulnerable to nephrotoxic insult. Many of drug-induced nephrotoxic effects observed in clinical trials remain undetected in preclinical testing models. Furthermore, although the kidney can regenerate after acute kidney injury, subclinical renal impairment sometimes persists, which increases the risk of chronic kidney disease. In this context, differentiation models based on stem cells offer a promising alternative to traditional in vitro systems, as they could provide more physiologically relevant, human-specific models for drug testing and disease modelling that align with the 3Rs (replace, reduce and refine) principles.
In this study, a one-step differentiation protocol adapted from Kandasamy et al. (2015) was used to differentiate two human-induced pluripotent stem cell (hiPSC) lines, b4 and UM51, into proximal tubule epithelial-like cells (PTELC). The differentiation efficiency was confirmed by qRT-PCR, western blot, immunocytochemistry, and also by albumin uptake assay. Dose-response experiments with well-established nephrotoxic substances like cisplatin, cyclosporin A (CsA) and hydrogen peroxide (H2O2), were performed on hiPSC, differentiating hiPSC and differentiated hiPSC. Further toxicity studies included cisplatin-induced oxidative stress response, analyzed by qRT-PCR, immunocytochemistry and glutathione (GSH) assay. Differentiated b4 and UM51 cells displayed typical morphological characteristics of PTEC, confirmed by the downregulation of stem cell markers and upregulation of PTEC-specific markers at both mRNA and protein levels. Functionally, PTELC showed enhanced albumin uptake capacity. Toxicity studies revealed consisting inhibitory concentration (IC) values with existing literature, with PTELC exhibiting reduced sensitivity to cisplatin, while CsA was similarly toxic across all cell stages of cell differentiation. Interestingly, differentiating cells were more sensitive to H2O2 than stem cells. Cisplatin had overall a more pronounced effect on cell proliferation than CsA. In oxidative stress response to cisplatin, PTELC exhibited a dose-dependent increase in glutathione peroxidase (GPX-1) and NADPH (quinone) dehydrogenase-1 (NQO-1) at low doses, while the antioxidant defence was overwhelmed at higher doses. In conclusion, our results suggest that the present hiPSC-derived PTELC model represents a promising 3R-compatible tool for regenerative medicine and drug toxicity studies. It could offer concrete advantages in predicting nephrotoxicity and improving preclinical drug screening. | |||||||
| Lizenz: |  Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz | |||||||
| Fachbereich / Einrichtung: | Medizinische Fakultät » Institute » Institut für Toxikologie | |||||||
| Dokument erstellt am: | 08.12.2025 | |||||||
| Dateien geändert am: | 08.12.2025 | |||||||
| Promotionsantrag am: | 08.05.2025 | |||||||
| Datum der Promotion: | 28.10.2025 |
