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Gene Editing in 3D Cultured Nephron Progenitor Cell Lines

  • Zhongwei LiEmail author
  • Toshikazu Araoka
  • Juan Carlos Izpisua BelmonteEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1926)

Abstract

During kidney development, Six2+ nephron progenitor cells (NPCs) generate nephrons, the functional units of the kidney. Isolating and expanding NPCs in vitro have enabled wide applications in both basic and translational research. Here we describe the methods to derive NPC lines from mouse embryonic kidneys in a 3D culture format as well as gene editing in the NPC lines.

Key words

Nephron progenitor cell (NPC) 3D culture NPSR culture medium Gene editing CRISPR-Cas9 

Notes

Acknowledgments

This work was supported by Fundacion Dr. Pedro Guillen, Universidad Catolica San Antonio de Murcia, G. Harold and Leila Y. Mathers Charitable Foundation, The Leona M. and Harry B. Helmsley Charitable Trust, The Moxie Foundation, and National Institute of Health (R21 AG055938).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Nephrology and Hypertension, Department of Medicine and USC/UKRO Kidney Research CenterKeck School of Medicine of the University of Southern CaliforniaLos AngelesUSA
  2. 2.Department of Stem Cell Biology and Regenerative MedicineKeck School of Medicine of the University of Southern CaliforniaLos AngelesUSA
  3. 3.Center for iPS Cell Research and Application (CiRA)Kyoto UniversityKyotoJapan
  4. 4.Gene Expression LaboratoryThe Salk Institute for Biological StudiesLa JollaUSA

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