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Formation of Mature Nephrons by Implantation of Human Pluripotent Stem Cell-Derived Progenitors into Mice

  • Ioannis BantounasEmail author
  • Edina Silajdžić
  • Adrian S. Woolf
  • Susan J. Kimber
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2067)

Abstract

In the future, stem cell-based technologies may be harnessed to replace conventional dialysis and transplantation in patients with diabetic nephropathy. Recently, there has been considerable effort to improve methods for the differentiation of human pluripotent stem cells (hPSCs) into kidney cells in culture. Here, we present a protocol for obtaining more advanced kidney structures than have currently been possible in vitro, including vascularized glomeruli and tubular elements. HPSCs are first differentiated in 2D culture to a kidney progenitor stage. These cells are then dissociated and injected subcutaneously into immunocompromised mice. Twelve weeks later, the cells have developed into mature kidney structures and are excised for further characterization. This method constitutes a significant improvement on protocols that involve either exclusively a 2D culture or placing the cells in 3D organoid culture at the air-liquid interface in vitro.

Key words

Stem cells hESC iPSC Kidney Nephron Organoid Differentiation Implant End-stage kidney disease 

Notes

Acknowledgments

We acknowledge grant support as follows: Research Councils UK/Medical Research Council (MRC) MR/K026739/1 UK Regenerative Medicine Platform Hub, Safety and Efficacy of Stem Cells; MRC MICA grant MR/M017344; Kidneys for Life pump priming grant KfL/1/15; Manchester Regenerative Medicine Network strategic funding initiative grant MARMN/1/17; and Kidney Research UK project grant JFS/RP/008/20160916.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Ioannis Bantounas
    • 1
    Email author
  • Edina Silajdžić
    • 1
  • Adrian S. Woolf
    • 1
    • 2
  • Susan J. Kimber
    • 1
  1. 1.Division of Cell Matrix Biology and Regenerative Medicine, Faculty of Biology, Medicine and HealthUniversity of Manchester, Manchester Academic Health Science CentreManchesterUK
  2. 2.Royal Manchester Children’s HospitalManchesterUK

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