The Directed Differentiation of Human iPS Cells into Kidney Podocytes

Bi Song1, Alexandra M. Smink1, Christina V. Jones1, Judy M. Callaghan2, Stephen D. Firth2, Claude A. Bernard1, Andrew L. Laslett3,4, Peter G. Kerr5, Sharon D. Ricardo1*

1 Monash Immunology and Stem Cell Laboratories (MISCL), Monash University, Clayton, Victoria, Australia, 2 Monash Micro Imaging, Monash University, Clayton, Victoria, Australia, 3 CSIRO Materials Science and Engineering, Clayton, Victoria, Australia, 4 Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia, 5 Department of Medicine, Monash University, Monash Medical Centre, Clayton, Victoria, Australia

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Abstract

The loss of glomerular podocytes is a key event in the progression of chronic kidney disease resulting in proteinuria and declining function. Podocytes are slow cycling cells that are considered terminally differentiated. Here we provide the first report of the directed differentiation of induced pluripotent stem (iPS) cells to generate kidney cells with podocyte features. The iPS-derived podocytes share a morphological phenotype analogous with cultured human podocytes. Following 10 days of directed differentiation, iPS podocytes had an up-regulated expression of mRNA and protein localization for podocyte markers including synaptopodin, nephrin and Wilm’s tumour protein (WT1), combined with a down-regulation of the stem cell marker OCT3/4. In contrast to human podocytes that become quiescent in culture, iPS-derived cells maintain a proliferative capacity suggestive of a more immature phenotype. The transduction of iPS podocytes with fluorescent labeled-talin that were immunostained with podocin showed a cytoplasmic contractile response to angiotensin II (AII). A permeability assay provided functional evidence of albumin uptake in the cytoplasm of iPS podocytes comparable to human podocytes. Moreover, labeled iPS-derived podocytes were found to integrate into reaggregated metanephric kidney explants where they incorporated into developing glomeruli and co-expressed WT1. This study establishes the differentiation of iPS cells to kidney podocytes that will be useful for screening new treatments, understanding podocyte pathogenesis, and offering possibilities for regenerative medicine.

Citation

Song B, Smink AM, Jones CV, Callaghan JM, Firth SD, et al. (2012) The Directed Differentiation of Human iPS Cells into Kidney Podocytes. PLoS ONE 7(9): e46453. doi:10.1371/journal.pone.0046453

Editor

Maria Pia Rastaldi, Fondazione IRCCS Ospedale Maggiore Policlinico & Fondazione D’Amico per la Ricerca sulle Malattie Renali, Italy

Received

July 21, 2012; Accepted: August 30, 2012; Published: September 28, 2012

Copyright

© Song et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding

This work was supported by the Alport Foundation of Australia and a Monash University Strategic Grant. CA Bernard is a recipient of an Erdi Fellowship in Neurological Diseases and funding from the Baker Foundation. Microscopy was performed with technical expertise from Monash Micro Imaging, Monash University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests

The authors have declared that no competing interests exist.

* E-mail: Sharon.Ricardo@monash.edu