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First published online 30 November 2005
doi: 10.1242/dev.02174
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1 Division of Stem Cell Regulation, The Institute of Medical Science, The
University of Tokyo, Tokyo 108-8639, Japan.
2 Department of Life Sciences (Biology), Graduate School of Arts and Sciences,
The University of Tokyo, Tokyo 153-8902, Japan.
3 ICORP, JST, Saitama 332-0012, Japan.
4 Division of Integrative Cell Biology, Institute of Molecular Embryology and
Genetics, Kumamoto University, Kumamoto 860-0811, Japan.
5 Institut für Molekularbiologie, Medizinische Hochschule Hannover, 30625
Hannover, Germany.
6 PRESTO, JST, Saitama 332-0012, Japan.
* Author for correspondence (e-mail: ryuichi{at}kaiju.medic.kumamoto-u.ac.jp)
Accepted 24 October 2005
Renal stem or progenitor cells with a multilineage differentiation potential remain to be isolated, and the differentiation mechanism of these cell types in kidney development or regeneration processes is unknown. In an attempt to resolve this issue, we set up an in vitro culture system using NIH3T3 cells stably expressing Wnt4 (3T3Wnt4) as a feeder layer, in which a single renal progenitor in the metanephric mesenchyme forms colonies consisting of several types of epithelial cells that exist in glomeruli and renal tubules. We found that only cells strongly expressing Sall1 (Sall1-GFPhigh cells), a zinc-finger nuclear factor essential for kidney development, form colonies, and that they reconstitute a three-dimensional kidney structure in an organ culture setting. We also found that Rac- and JNK-dependent planar cell polarity (PCP) pathways downstream of Wnt4 positively regulate the colony size, and that the JNK pathway is also involved in mesenchymal-to-epithelial transformation of colony-forming progenitors. Thus our colony-forming assay, which identifies multipotent progenitors in the embryonic mouse kidney, can be used for examining mechanisms of renal progenitor differentiation.
Key words: Progenitor, Kidney, Colony-forming assay, Sall1, Wnt, PCP, JNK, Rho, Mouse
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