Large-Scale Production of Kidney Organoids from Human Pluripotent Stem Cells

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 Jul 9

PMID 37423983

Authors

Veronika Sander

Aneta Przepiorski

Neil A Hukriede

Alan J Davidson

Affiliations

Soon will be listed here.

Abstract

Kidney organoids differentiated from human pluripotent stem cells (hPSC) have advanced the study of kidney diseases by providing an in vitro system that outperforms traditional monolayer cell culture and complements animal models. This chapter describes a simple two-stage protocol that generates kidney organoids in suspension culture in less than 2 weeks. In the first stage, hPSC colonies are differentiated into nephrogenic mesoderm. In the second stage of the protocol, renal cell lineages develop and self-organize into kidney organoids that contain fetal-like nephrons with proximal and distal tubule segmentation. A single assay generates up to 1000 organoids, thereby providing a rapid and cost-efficient method for the bulk production of human kidney tissue. Applications include the study of fetal kidney development, genetic disease modelling, nephrotoxicity screening, and drug development.

Citing Articles

The biogenesis of potassium transporters: implications of disease-associated mutations.

Kok M, Brodsky J Crit Rev Biochem Mol Biol. 2024; 59(3-4):154-198.

PMID: 38946646 PMC: 11444911. DOI: 10.1080/10409238.2024.2369986.

References

Kim J, Koo B, Knoblich J . Human organoids: model systems for human biology and medicine. Nat Rev Mol Cell Biol. 2020; 21(10):571-584. PMC: 7339799. DOI: 10.1038/s41580-020-0259-3. View

Taguchi A, Kaku Y, Ohmori T, Sharmin S, Ogawa M, Sasaki H . Redefining the in vivo origin of metanephric nephron progenitors enables generation of complex kidney structures from pluripotent stem cells. Cell Stem Cell. 2013; 14(1):53-67. DOI: 10.1016/j.stem.2013.11.010. View

Takasato M, Er P, Chiu H, Maier B, Baillie G, Ferguson C . Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis. Nature. 2015; 526(7574):564-8. DOI: 10.1038/nature15695. View

Morizane R, Lam A, Freedman B, Kishi S, Valerius M, Bonventre J . Nephron organoids derived from human pluripotent stem cells model kidney development and injury. Nat Biotechnol. 2015; 33(11):1193-200. PMC: 4747858. DOI: 10.1038/nbt.3392. View

Freedman B, Brooks C, Lam A, Fu H, Morizane R, Agrawal V . Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids. Nat Commun. 2015; 6:8715. PMC: 4620584. DOI: 10.1038/ncomms9715. View

Low J, Li P, Chew E, Zhou B, Suzuki K, Zhang T . Generation of Human PSC-Derived Kidney Organoids with Patterned Nephron Segments and a De Novo Vascular Network. Cell Stem Cell. 2019; 25(3):373-387.e9. PMC: 6731150. DOI: 10.1016/j.stem.2019.06.009. View

Uchimura K, Wu H, Yoshimura Y, Humphreys B . Human Pluripotent Stem Cell-Derived Kidney Organoids with Improved Collecting Duct Maturation and Injury Modeling. Cell Rep. 2020; 33(11):108514. PMC: 10122187. DOI: 10.1016/j.celrep.2020.108514. View

Combes A, Zappia L, Er P, Oshlack A, Little M . Single-cell analysis reveals congruence between kidney organoids and human fetal kidney. Genome Med. 2019; 11(1):3. PMC: 6345028. DOI: 10.1186/s13073-019-0615-0. View

Wu H, Uchimura K, Donnelly E, Kirita Y, Morris S, Humphreys B . Comparative Analysis and Refinement of Human PSC-Derived Kidney Organoid Differentiation with Single-Cell Transcriptomics. Cell Stem Cell. 2018; 23(6):869-881.e8. PMC: 6324730. DOI: 10.1016/j.stem.2018.10.010. View

10.

Czerniecki S, Cruz N, Harder J, Menon R, Annis J, Otto E . High-Throughput Screening Enhances Kidney Organoid Differentiation from Human Pluripotent Stem Cells and Enables Automated Multidimensional Phenotyping. Cell Stem Cell. 2018; 22(6):929-940.e4. PMC: 5984728. DOI: 10.1016/j.stem.2018.04.022. View

11.

Digby J, Vanichapol T, Przepiorski A, Davidson A, Sander V . Evaluation of cisplatin-induced injury in human kidney organoids. Am J Physiol Renal Physiol. 2020; 318(4):F971-F978. PMC: 7395477. DOI: 10.1152/ajprenal.00597.2019. View

12.

Hale L, Howden S, Phipson B, Lonsdale A, Er P, Ghobrial I . 3D organoid-derived human glomeruli for personalised podocyte disease modelling and drug screening. Nat Commun. 2018; 9(1):5167. PMC: 6279764. DOI: 10.1038/s41467-018-07594-z. View

13.

Lawlor K, Vanslambrouck J, Higgins J, Chambon A, Bishard K, Arndt D . Cellular extrusion bioprinting improves kidney organoid reproducibility and conformation. Nat Mater. 2020; 20(2):260-271. PMC: 7855371. DOI: 10.1038/s41563-020-00853-9. View

14.

Lemos D, McMurdo M, Karaca G, Wilflingseder J, Leaf I, Gupta N . Interleukin-1 Activates a MYC-Dependent Metabolic Switch in Kidney Stromal Cells Necessary for Progressive Tubulointerstitial Fibrosis. J Am Soc Nephrol. 2018; 29(6):1690-1705. PMC: 6054344. DOI: 10.1681/ASN.2017121283. View

15.

Przepiorski A, Vanichapol T, Espiritu E, Crunk A, Parasky E, McDaniels M . Modeling oxidative injury response in human kidney organoids. Stem Cell Res Ther. 2022; 13(1):76. PMC: 8862571. DOI: 10.1186/s13287-022-02752-z. View

16.

Forbes T, Howden S, Lawlor K, Phipson B, Maksimovic J, Hale L . Patient-iPSC-Derived Kidney Organoids Show Functional Validation of a Ciliopathic Renal Phenotype and Reveal Underlying Pathogenetic Mechanisms. Am J Hum Genet. 2018; 102(5):816-831. PMC: 5986969. DOI: 10.1016/j.ajhg.2018.03.014. View

17.

Hollywood J, Przepiorski A, DSouza R, Sreebhavan S, Wolvetang E, Harrison P . Use of Human Induced Pluripotent Stem Cells and Kidney Organoids To Develop a Cysteamine/mTOR Inhibition Combination Therapy for Cystinosis. J Am Soc Nephrol. 2020; 31(5):962-982. PMC: 7217405. DOI: 10.1681/ASN.2019070712. View

18.

Sander V, Przepiorski A, Crunk A, Hukriede N, Holm T, Davidson A . Protocol for Large-Scale Production of Kidney Organoids from Human Pluripotent Stem Cells. STAR Protoc. 2020; 1(3):100150. PMC: 7757290. DOI: 10.1016/j.xpro.2020.100150. View

19.

Orlova V, van den Hil F, Petrus-Reurer S, Drabsch Y, Ten Dijke P, Mummery C . Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells. Nat Protoc. 2014; 9(6):1514-31. DOI: 10.1038/nprot.2014.102. View

20.

Ng E, Davis R, Stanley E, Elefanty A . A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies. Nat Protoc. 2008; 3(5):768-76. DOI: 10.1038/nprot.2008.42. View