Advancing Kidney Organoid Development for Regenerative Nephrology: Enhancing Differentiation and Functionality Using hiPSCs and Bioprinting

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https://storage.unitedwebnetwork.com/files/1099/dd88e4e0c2f2eeacde30a3eb62943ba1.pdf

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Advancing Kidney Organoid Development for Regenerative Nephrology: Enhancing Differentiation and Functionality Using hiPSCs and Bioprinting

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Co-author 1

Carlos Mota c.mota@maastrichtuniversity.nl Maastricht University Department of Complex Tissue Regeneration Maastricht Netherlands *

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Introduction

Chronic kidney disease (CKD) affects approximately 10% of the global population and frequently progresses to end-stage renal disease, where dialysis or transplantation are the only available treatments. Human induced pluripotent stem cells (hiPSCs) offer a promising platform for regenerative nephrology, enabling the development of kidney organoids for disease modeling, drug screening, and potential therapeutic applications. However, current differentiation protocols often yield immature and structurally limited organoids, and manual production introduces variability that hampers scalability.

Methods

To improve differentiation efficiency, hiPSCs were treated with low-dose dimethyl sulfoxide (DMSO, 1–2%) prior to initiating kidney organoid formation using a stepwise 2D monolayer protocol. Changes in pluripotency gene expression, colony morphology, and epigenetic markers were assessed. In parallel, hiPSC-derived metanephric mesenchyme and ureteric bud progenitors were generated and bioprinted using a microfluidic system that produced core-shell filaments—gelatin in the core and alginate in the shell. These constructs were cultured in a growth factor-enriched medium for two weeks.

Results

DMSO treatment modulated the expression of key pluripotency transcription factors and enhanced the expression of SIX2, a nephron progenitor marker, by day 9 of differentiation. Bioprinted constructs showed renal vesicle formation within one week and developed into functional kidney organoids by week two. These organoids responded to nephrotoxic exposure (doxorubicin), indicating physiological relevance and functional maturation.

Conclusion

Combining chemical modulation with automated bioprinting significantly improves the differentiation and functional development of kidney organoids from hiPSCs. This integrated approach enhances reproducibility, scalability, and physiological relevance, offering a robust platform for nephrology research and future regenerative therapies.

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