A Simple Bioreactor-Based Method to Generate Kidney Organoids from Pluripotent Stem Cells

• Published in: Stem cell reports Vol. 11; no. 2; pp. 470 - 484
• Main Authors: Przepiorski, Aneta, Sander, Veronika, Tran, Tracy, Hollywood, Jennifer A., Sorrenson, Brie, Shih, Jen-Hsing, Wolvetang, Ernst J., McMahon, Andrew P., Holm, Teresa M., Davidson, Alan J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 14.08.2018; Elsevier
• Subjects: 3D culture; Biomarkers; bioreactor; Bioreactors; Cell Culture Techniques; Cell Differentiation; CRISPR/Cas9; embryoid body; fetal human kidney; Fibrosis; Gene Knockout Techniques; Hepatocyte Nuclear Factor 1-beta - genetics; HNF1B; Humans; Induced Pluripotent Stem Cells - cytology; iPSC; Kidney - cytology; Kidney - embryology; kidney organoid; Nephrons - cytology; Organoids - cytology; Pluripotent Stem Cells - cytology; renal development; 3D culture; fetal human kidney; CRISPR/Cas9; renal development; iPSC; HNF1B; fibrosis; kidney organoid; bioreactor; embryoid body
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2018.06.018

Kidney organoids made from pluripotent stem cells have the potential to revolutionize how kidney development, disease, and injury are studied. Current protocols are technically complex, suffer from poor reproducibility, and have high reagent costs that restrict scalability. To overcome some of these issues, we have established a simple, inexpensive, and robust method to grow kidney organoids in bulk from human induced pluripotent stem cells. Our organoids develop tubular structures by day 8 and show optimal tissue morphology at day 14. A comparison with fetal human kidneys suggests that day-14 organoid tissue most closely resembles late capillary loop stage nephrons. We show that deletion of HNF1B, a transcription factor linked to congenital kidney defects, interferes with tubulogenesis, validating our experimental system for studying renal developmental biology. Taken together, our protocol provides a fast, efficient, and cost-effective method for generating large quantities of human fetal kidney tissue, enabling the study of normal and aberrant kidney development. [Display omitted] •Technically simple and cost-efficient protocol for kidney organoid generation•Tubular organoids are obtained rapidly, with high efficiency, yield, and robustness•Organoids contain nephrons that correspond to human fetal nephrons•The applicability to model congenital kidney defects is presented In this study, Przepiorski et al. present a technically simple and robust protocol to derive kidney organoids from human induced pluripotent stem cells. This technique is inexpensive and efficient, allowing kidney organoids to be generated in bulk. Organoids are obtained rapidly (within 8 days) and, based on their segmentation pattern, correspond to “late capillary loop” stage fetal human nephrons.