Gene Editing in 3D Cultured Nephron Progenitor Cell Lines

Zhongwei Li; Toshikazu Araoka; Juan Carlos Izpisua Belmonte

doi:10.1007/978-1-4939-9021-4_13

Gene Editing in 3D Cultured Nephron Progenitor Cell Lines

Methods Mol Biol. 2019:1926:151-159. doi: 10.1007/978-1-4939-9021-4_13.

Authors

Zhongwei Li^{1

2}, Toshikazu Araoka³, Juan Carlos Izpisua Belmonte⁴

Affiliations

¹ Division of Nephrology and Hypertension, Department of Medicine and USC/UKRO Kidney Research Center, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA. zhongwei.li@med.usc.edu.
² Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA. zhongwei.li@med.usc.edu.
³ Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
⁴ Gene Expression Laboratory, The Salk Institute for Biological Studies, La Jolla, CA, USA. belmonte@salk.edu.

PMID: 30742270
DOI: 10.1007/978-1-4939-9021-4_13

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.

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

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Cells, Cultured
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Gene Editing / methods
Mice
Nephrons / cytology*
Stem Cells / cytology*

Grants and funding

R21 AG055938/AG/NIA NIH HHS/United States