Parallel generation of easily selectable multiple nephronal cell types from human pluripotent stem cells
Human pluripotent stem cells (hPSCs) provide a source for the generation of defined kidney cells and renal organoids applicable in regenerative medicine, disease modeling, and drug screening. These applications require the provision of hPSC-derived renal cells by reproducible, scalable, and efficient methods. We established a chemically defined protocol by application of Activin A, BMP4, and Retinoic acid followed by GDNF, which steered hPSCs to the renal lineage and resulted in populations of SIX2+/CITED1+ metanephric mesenchyme- (MM) and of HOXB7+/GRHL2+ ureteric bud (UB)-like cells already by 6 days. Transcriptome analysis corroborated that the PSC-derived cell types at day 8 resemble their renal vesicle and ureteric epithelial counterpart in vivo, forming tubular and glomerular renal cells 6 days later. We demonstrate that starting from hPSCs, our in vitro protocol generates a pool of nephrogenic progenitors at the renal vesicle stage, which can be further directed into specialized nephronal cell types including mesangial-, proximal tubular-, distal tubular, collecting duct epithelial cells, and podocyte precursors after 14 days. This simple and rapid method to produce renal cells from a common precursor pool in 2D culture provides the basis for scaled-up production of tailored renal cell types, which are applicable for drug testing or cell-based regenerative therapies.
KeywordsHPSCS Renal progenitors Differentiation Tubules Renal vesicle Nephron
This work was supported by Berlin-Brandenburg School for Regenerative Therapies and the German Federal Ministry of Education and Research (VIP, FKZ 03V0396) and (FKZ 031A303B).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
- 10.Sharmin S, Taguchi A, Kaku Y, Yoshimura Y, Ohmori T, Sakuma T, Mukoyama M, Yamamoto T, Kurihara H, Nishinakamura R (2015) Human induced pluripotent stem cell-derived podocytes mature into vascularized glomeruli upon experimental transplantation. J Am Soc Nephrol 27(6):1778–1791CrossRefGoogle Scholar
- 12.Sumi T, Tsuneyoshi N, Nakatsuji N, Suemori H (2008) Defining early lineage specification of human embryonic stem cells by the orchestrated balance of canonical Wnt/beta-catenin, Activin/Nodal and BMP signaling. Dev Camb Engl. 135(17):2969–2979Google Scholar
- 15.Majumdar A, Vainio S, Kispert A, McMahon J, McMahon AP (2003) Wnt11 and Ret/Gdnf pathways cooperate in regulating ureteric branching during metanephric kidney development. Dev Camb Engl 130(14):3175–3185Google Scholar
- 17.Xu P-X, Zheng W, Huang L, Maire P, Laclef C, Silvius D (2003) Six1 is required for the early organogenesis of mammalian kidney. Dev Camb Engl 130(14):3085–3094Google Scholar
- 18.Kanda S, Tanigawa S, Ohmori T, Taguchi A, Kudo K, Suzuki Y, Sato Y, Hino S, Sander M, Perantoni AO, Sugano S, Nakao M, Nishinakamura R (2014) Sall1 maintains nephron progenitors and nascent nephrons by acting as both an activator and a repressor. J Am Soc Nephrol 25(11):2584–2595CrossRefGoogle Scholar
- 29.Kandasamy K, Chuah JKC, Su R, Huang P, Eng KG, Xiong S, Li Y, Chia CS, Loo LH, Zink D (2015) Prediction of drug-induced nephrotoxicity and injury mechanisms with human induced pluripotent stem cell-derived cells and machine learning methods. Sci Rep 5:12337. https://doi.org/10.1038/srep12337 CrossRefPubMedPubMedCentralGoogle Scholar
- 34.Yamanaka S, Yokoo T, Yamanaka S, Yokoo T (2015) Current bioengineering methods for whole kidney regeneration, current bioengineering methods for whole kidney regeneration. Stem Cells Int Stem Cells Int. 2015:e724047Google Scholar
- 40.R Core Team (2014) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, AustriaGoogle Scholar