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Parallel generation of easily selectable multiple nephronal cell types from human pluripotent stem cells

  • Krithika Hariharan
  • Harald Stachelscheid
  • Bella Rossbach
  • Su-Jun Oh
  • Nancy Mah
  • Kai Schmidt-Ott
  • Andreas Kurtz
  • Petra Reinke
Original Article
  • 6 Downloads

Abstract

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.

Keywords

HPSCS Renal progenitors Differentiation Tubules Renal vesicle Nephron 

Notes

Acknowledgements

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.

Supplementary material

18_2018_2929_MOESM1_ESM.docx (2.9 mb)
Supplementary material 1 (DOCX 3006 kb)

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Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  • Krithika Hariharan
    • 1
  • Harald Stachelscheid
    • 1
    • 2
  • Bella Rossbach
    • 1
  • Su-Jun Oh
    • 1
  • Nancy Mah
    • 1
  • Kai Schmidt-Ott
    • 3
    • 4
  • Andreas Kurtz
    • 1
  • Petra Reinke
    • 1
    • 3
  1. 1.Berlin-Brandenburg Center for Regenerative Therapies (BCRT)Charité-Universitätsmedizin BerlinBerlinGermany
  2. 2.Berlin Institute of Health (BIH), Stem Cell CoreBerlinGermany
  3. 3.Department of Nephrology and Intensive CareCharité University Medicine BerlinBerlinGermany
  4. 4.Max Delbrueck Center for Molecular Medicine in the Helmholtz AssociationBerlinGermany

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