Systematic production of human kidney organoids for transplantation in porcine kidneys during ex vivo machine perfusion

Organoids derived from human pluripotent stem (hPS) cells hold promise for therapeutic purposes. However, technological advances to overcome their massive production while ensuring differentiation fidelity are still lacking. Here we report a procedure sustaining the derivation of kidney organoids from hPS cells (hPSC-kidney organoids) using a scalable, reproducible and affordable approach that allows hPSC-kidney organoid differentiation into different renal cell types. Using single-cell RNA sequencing, confocal image analysis, metabolic assays and CRISPR-Cas9 engineering for generation of fluorescent reporters, we show that hPSC-kidney organoids exhibit transcriptional variety and cellular composition following cell-to-cell contact. We infuse human kidney organoids into ex vivo porcine kidneys using normothermic machine perfusion, and demonstrate in vivo engraftment of hPSC-kidney organoids. We further evaluate the immune response, confirming the feasibility and viability of the procedure. We identify cells of human origin after normothermic machine perfusion and in vivo transplantation by means of in situ hybridization, immunohistochemistry, confocal microscopy, image analysis and quantification, in vivo imaging, and flow cytometry. This work provides a foundation for using hPSC-kidney organoids for ex vivo cell-based therapies in clinical trials.

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Publication Details

Journal
Nature Biomedical Engineering
Published
2025-10-31
DOI
https://doi.org/10.1038/s41551-025-01542-1
Citations
6
Primary Topic
Renal and related cancers
Type
article
Field-Weighted Citation Impact
4.84
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article

Systematic production of human kidney organoids for transplantation in porcine kidneys during ex vivo machine perfusion

Gaia Amato, Henri G. D. Leuvenink, Asier Ullate‐Agote, Juan R. Rodríguez-Madoz et al.
6 citations
Nature Biomedical Engineering
Renal and related cancers
4.84
article

Systematic production of human kidney organoids for transplantation in porcine kidneys during ex vivo machine perfusion

Gaia Amato, Henri G. D. Leuvenink, Asier Ullate‐Agote, Juan R. Rodríguez-Madoz, Jordi Ochando, Carlos J. Aranda, Felipe Prósper, Cyril Moers, Margarita González‐Martín, Beatriz Domínguez‐Gil, Maarten Naesens, Cristóbal Belda-Iniesta, Núria Montserrat, Roger Oria, Daniel Lozano‐Ojalvo, Daniel Moya‐Rull, Elena Garreta, Javier Rodríguez-Rivera, Carolina Tarantino, Aleix Gavaldà‐Navarro, Idoia Lucía Selfa, Alejandro Montero Salinas, Priyanka Koshy, Federico Gonzãlez, Tim L. Hamelink, Patrícia Martin, Amaia Vilas‐Zornoza, Andrés Marco, Merel B. F. Pool, Antón Fernández García, Alberto Centeno, Inmaculada Márquez-Leiva
article en
6 citations

Abstract

Organoids derived from human pluripotent stem (hPS) cells hold promise for therapeutic purposes. However, technological advances to overcome their massive production while ensuring differentiation fidelity are still lacking. Here we report a procedure sustaining the derivation of kidney organoids from hPS cells (hPSC-kidney organoids) using a scalable, reproducible and affordable approach that allows hPSC-kidney organoid differentiation into different renal cell types. Using single-cell RNA sequencing, confocal image analysis, metabolic assays and CRISPR-Cas9 engineering for generation of fluorescent reporters, we show that hPSC-kidney organoids exhibit transcriptional variety and cellular composition following cell-to-cell contact. We infuse human kidney organoids into ex vivo porcine kidneys using normothermic machine perfusion, and demonstrate in vivo engraftment of hPSC-kidney organoids. We further evaluate the immune response, confirming the feasibility and viability of the procedure. We identify cells of human origin after normothermic machine perfusion and in vivo transplantation by means of in situ hybridization, immunohistochemistry, confocal microscopy, image analysis and quantification, in vivo imaging, and flow cytometry. This work provides a foundation for using hPSC-kidney organoids for ex vivo cell-based therapies in clinical trials.

Nature Biomedical Engineering
Institució Catalana de Recerca i Estudis Avançats (ES), University Medical Center Groningen (NL), University of Groningen (NL), University of California, San Francisco (US), California Institute for Regenerative Medicine (US), Organización Nacional de Trasplantes (ES), Instituto de Salud Carlos III (ES), Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (ES), Centro de Investigación Biomédica en Red (ES), HM Hospitales (ES), Instituto de Investigación en Ciencias de la Alimentación (ES), Institute for Bioengineering of Catalonia (ES), Andalusian Centre for Nanomedicine and Biotechnology (ES), Centro de Investigación Biomédica en Red de Cáncer (ES), Instituto de Investigación Biomédica de A Coruña (ES), Instituto de Investigación Biomédica de Málaga (ES), Universidad Autónoma de Madrid (ES), Universitat de Barcelona (ES), Universidad de Navarra (ES), KU Leuven (BE)
Openalex Percentile: Top 5%
Renal and related cancers
4.84
DOIOpen Access PDFOpenAlex
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