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Stem Cell Reports. 2017 Feb 14;8(2):401-416. doi: 10.1016/j.stemcr.2016.12.011. Epub 2017 Jan 12.

In Vitro Propagation and Branching Morphogenesis from Single Ureteric Bud Cells.

Author information

1
Medical and Research Services, Greater Los Angeles Veterans Affairs Healthcare System at Sepulveda, North Hills, CA 91343, USA; Department of Medicine, University of California at Los Angeles, David Geffen School of Medicine, Los Angeles, CA 90095, USA. Electronic address: shunsukeyuri@ucla.edu.
2
Medical and Research Services, Greater Los Angeles Veterans Affairs Healthcare System at Sepulveda, North Hills, CA 91343, USA; Department of Medicine, University of California at Los Angeles, David Geffen School of Medicine, Los Angeles, CA 90095, USA.
3
Medical and Research Services, Greater Los Angeles Veterans Affairs Healthcare System at Sepulveda, North Hills, CA 91343, USA; Department of Medicine, University of California at Los Angeles, David Geffen School of Medicine, Los Angeles, CA 90095, USA. Electronic address: nori@ucla.edu.

Abstract

A method to maintain and rebuild ureteric bud (UB)-like structures from UB cells in vitro could provide a useful tool for kidney regeneration. We aimed in our present study to establish a serum-free culture system that enables the expansion of UB progenitor cells, i.e., UB tip cells, and reconstruction of UB-like structures. We found that fibroblast growth factors or retinoic acid (RA) was sufficient for the survival of UB cells in serum-free condition, while the proliferation and maintenance of UB tip cells required glial cell-derived neurotrophic factor together with signaling from either WNT-β-catenin pathway or RA. The activation of WNT-β-catenin signaling in UB cells by endogenous WNT proteins required R-spondins. Together with Rho kinase inhibitor, our culture system facilitated the expansion of UB tip cells to form UB-like structures from dispersed single cells. The UB-like structures thus formed retained the original UB characteristics and integrated into the native embryonic kidneys.

KEYWORDS:

R-spondin; branching morphogenesis; fibroblast growth factor; kidney progenitor cells; retinoic acid; ureteric bud

PMID:
28089670
PMCID:
PMC5311471
DOI:
10.1016/j.stemcr.2016.12.011
[Indexed for MEDLINE]
Free PMC Article

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