Article

Mature induced-pluripotent-stem-cell-derived human podocytes reconstitute kidney glomerular-capillary-wall function on a chip

  • Nature Biomedical Engineering 1, Article number: 0069 (2017)
  • doi:10.1038/s41551-017-0069
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Abstract

An in vitro model of the human kidney glomerulus—the major site of blood filtration—could facilitate drug discovery and illuminate kidney-disease mechanisms. Microfluidic organ-on-a-chip technology has been used to model the human proximal tubule, yet a kidney-glomerulus-on-a-chip has not been possible because of the lack of functional human podocytes—the cells that regulate selective permeability in the glomerulus. Here, we demonstrate an efficient (over 90%) and chemically defined method for directing the differentiation of human induced pluripotent stem (hiPS) cells into podocytes that express markers for a mature phenotype (nephrin+, WT1+, podocin+, PAX2) and that exhibit primary and secondary foot processes. We also show that the hiPS-cell-derived podocytes produce glomerular basement-membrane collagen and recapitulate the natural tissue–tissue interface of the glomerulus, as well as the differential clearance of albumin and inulin, when co-cultured with human glomerular endothelial cells in an organ-on-a-chip microfluidic device. The glomerulus-on-a-chip also mimics adriamycin-induced albuminuria and podocyte injury. This in vitro model of human glomerular function with mature human podocytes may facilitate drug development and personalized-medicine applications.

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Acknowledgements

This work was supported by the Defense Advanced Research Projects Agency under Cooperative Agreement Number W911NF-12-2-0036 and the Wyss Institute for Biologically Inspired Engineering at Harvard University. S.M. was supported by a Dean’s Postdoctoral Fellowship from Harvard Medical School, a UNCF-Merck Postdoctoral Fellowship, a Postdoctoral Enrichment Program Award from the Burroughs Wellcome Fund and an NIH/NIDDK Nephrology Training Grant (4T32DK007199-39). We thank the Wyss Institute Microfabrication team for organ-chip production; A. P. Mehr, K. Jang, A. Bahinski and R. Prantil-Baun for helpful discussions; E. Jiang, Y. Torisawa, E.I. Qendro and S. Lightbown for technical assistance, and R. Luna for helpful comments on the manuscript.

Author information

Affiliations

  1. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.

    • Samira Musah
    • , Thomas C. Ferrante
    • , Sauveur S. F. Jeanty
    • , Mariko Hirano-Kobayashi
    • , Kristen Roberts
    • , Seyoon Chung
    • , Richard Novak
    • , Miles Ingram
    • , Tohid Fatanat-Didar
    • , Sandeep Koshy
    • , James C. Weaver
    • , George M. Church
    •  & Donald E. Ingber
  2. Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.

    • Samira Musah
    •  & George M. Church
  3. Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts 02138, USA.

    • Samira Musah
    • , George M. Church
    •  & Donald E. Ingber
  4. Vascular Biology Program, Children’s Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.

    • Akiko Mammoto
    • , Mariko Hirano-Kobayashi
    • , Tadanori Mammoto
    •  & Donald E. Ingber
  5. Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02139, USA.

    • Donald E. Ingber

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Contributions

S.M., G.M.C. and D.E.I. conceived the strategy for this study; S.M. designed and performed experiments; S.M. and D.E.I. wrote the manuscript; T.M., T.C.F. and S.S.F.J. helped with the analysis of microscopy data; A.M. performed qPCR analysis; scanning electron microscopy analysis was performed by K.R., T.F.-D., S.K. and J.C.W.; M.H.-K. performed western blot experiments and analysed the data; S.C. performed LDH-release assay; S.M. and S.S.F.J. performed drug toxicity studies and analysed the data; R.N. and M.I. designed the microfluidic chips and built the programmable vacuum regulators. All authors discussed the results and commented on the manuscript.

Competing interests

D.E.I. and S.M. are authors on a pending patent for methods for the generation of kidney glomerular podocytes from pluripotent stem cells (US patent application 14/950859). D.E.I. is a founder of Emulate, Inc., holds equity in it, and chairs its scientific advisory board.

Corresponding author

Correspondence to Donald E. Ingber.

Supplementary information

PDF files

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    Supplementary Information

    Supplementary text, tables and video captions.

Videos

  1. 1.

    Supplementary Video 1

    Human iPS-derived podocytes and glomerular endothelial cells cultured in an organ-on-a-chip microfluidic device under fluid flow without strain.

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    Supplementary Video 2

    Human iPS-derived podocytes and glomerular endothelial cells cultured in an organ-on-a-chip microfluidic device under both fluid flow and mechanical strain.