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Stem Cell Res. 2016 Jul;17(1):130-9. doi: 10.1016/j.scr.2016.06.001. Epub 2016 Jun 3.

Generation of functional podocytes from human induced pluripotent stem cells.

Author information

1
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy; Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
2
Roche Pharma Research & Early Development, Roche Innovation Center Basel, 4070 Basel, Switzerland.
3
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy.
4
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy; Unit of Nephrology and Dialysis, Azienda Socio Sanitaria Territoriale (ASST) Papa Giovanni XXIII, 24127 Bergamo, Italy; Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy.
5
IRCCS - Istituto di Ricerche Farmacologiche Mario Negri, Centro Anna Maria Astori, Science and Technology Park Kilometro Rosso, 24126 Bergamo, Italy. Electronic address: ariela.benigni@marionegri.it.

Abstract

Generating human podocytes in vitro could offer a unique opportunity to study human diseases. Here, we describe a simple and efficient protocol for obtaining functional podocytes in vitro from human induced pluripotent stem cells. Cells were exposed to a three-step protocol, which induced their differentiation into intermediate mesoderm, then into nephron progenitors and, finally, into mature podocytes. After differentiation, cells expressed the main podocyte markers, such as synaptopodin, WT1, α-Actinin-4, P-cadherin and nephrin at the protein and mRNA level, and showed the low proliferation rate typical of mature podocytes. Exposure to Angiotensin II significantly decreased the expression of podocyte genes and cells underwent cytoskeleton rearrangement. Cells were able to internalize albumin and self-assembled into chimeric 3D structures in combination with dissociated embryonic mouse kidney cells. Overall, these findings demonstrate the establishment of a robust protocol that, mimicking developmental stages, makes it possible to derive functional podocytes in vitro.

KEYWORDS:

Differentiation; Induced pluripotent stem cells; Nephron progenitors; Podocytes

PMID:
27299470
PMCID:
PMC5009184
DOI:
10.1016/j.scr.2016.06.001
[Indexed for MEDLINE]
Free PMC Article

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