Making a Kidney Organoid Using the Directed Differentiation of Human Pluripotent Stem Cells

Part of the Methods in Molecular Biology book series (MIMB, volume 1597)

Abstract

An organoid can be defined as a three-dimensional organ-like structure formed from organ-specific progenitor cells. Organ progenitor cells were empirically found to self-organize three-dimensional tissues when they were aggregated and cultivated in vitro. While this nature power of progenitor cells has an amazing potential to recreate artificial organs in vitro, there had been difficulty to apply this technology to human organs due to the inaccessibility to human progenitor cells until human-induced pluripotent stem cell (hiPSC) was invented by Takahashi and Yamanaka in 2007. As embryonic stem cells do, hiPSCs also have pluripotency to give rise to any organs/tissues cell types, including the kidney, via directed differentiation. Here, we provide a detailed protocol for generating kidney organoids using human pluripotent stem cells. The protocol differentiates human pluripotent stem cells into the posterior primitive streak. This is followed by the simultaneous induction of posterior and anterior intermediate mesoderm that are subsequently aggregated and undergo self-organization into the kidney organoid. Such kidney organoids are comprised of all anticipated kidney cell types including nephrons segmented into the glomerulus, proximal tubule, loop of Henle, and distal tubule as well as the collecting duct, endothelial network, and renal interstitium.

Key words

Kidney organoid Kidney development Directed differentiation Human pluripotent stem cells Early embryogenesis Primitive streak Intermediate mesoderm Metanephric mesenchyme Ureteric bud 

Notes

Acknowledgments

M.L. is a National Health and Medical Research Council Senior Principal Research Fellow. This work was supported by the NHMRC and the Australian Research Council. M.C.R.I. is supported by the Victorian Government’s Operational Infrastructure Support Program.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Murdoch Children’s Research InstituteParkvilleAustralia
  2. 2.RIKEN Center for Developmental BiologyKobeJapan
  3. 3.Department of PediatricsUniversity of MelbourneParkvilleAustralia

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