A Simple Bioreactor-Based Method to Generate Kidney Organoids from Pluripotent Stem Cells
Creators
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1.
University of Auckland
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2.
University of Southern California
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3.
University of Queensland
Abstract
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.
Copyright and License
© 2018 The Authors. Creatives Commons - Attribution-NonCommercial-NoDerivatives 4.0 International.
Acknowledgement
We thank Adrian Turner, Jacqui Ross, and Satya Amirapu for help with histology and microscopy, Paula Lewis for critical reading of the manuscript, and Peter Shepherd for providing resources to facilitate the project. Work in A.J.D.’s laboratory was supported by the Health Research Council of New Zealand (17/425), Auckland Medical Research Foundation (1116018), Cystinosis Research Foundation USA, Cystinosis Research Ireland Foundation (MRCG-2014-8) Maurice Wilkins Center, and Valrae Collins Philanthropic support for A.P. Work in A.P.M.’s laboratory was supported by the NIH (DK054364) and California Institute of Regenerative Medicine (LA1-06536).
Contributions
A.P., T.M.H., and A.J.D. conceptualized the study; A.P. and V.S. designed the experiments; A.P. established the protocol; A.P. and V.S. conducted organoid culture, processing, and analysis; V.S. generated the HNF1B knockout; T.T. analyzed human fetal kidney tissue; A.P., J.A.H., J.-H.S., B.S., and T.M.H. generated the MANZ-2-2 iPSC line; E.J.W. generated the C32 iPSC line; A.P.M. and T.M.H. co-supervised the study; A.P., V.S., and A.J.D. wrote the manuscript; A.J.D. supervised the study; V.S., A.P.M., and A.J.D. acquired funding.
Supplemental Material
Document S1. Supplemental Experimental Procedures, Figures S1–S7, and Tables S1 and S2.
Document S2. Article plus Supplemental Information.
Files
1-s2.0-S2213671118302819-main.pdf
Additional details
Related works
- Has version
- Discussion Paper: 10.1101/237644 (DOI)
- Is referenced by
- Journal Article: 10.1016/j.xpro.2020.100150 (DOI)
Funding
- Health Research Council of New Zealand
- 17/425
- Auckland Medical Research Foundation
- 1116018
- Cystinosis Research Foundation
- Cystinosis Ireland
- MRCG-2014-8
- Maurice Wilkins Centre
- National Institutes of Health
- DK054364
- California Institute for Regenerative Medicine
- LA1-06536
Dates
- Accepted
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2018-06-18Accepted
- Available
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2018-07-19Published online
- Available
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2018-08-14Version of record