A Modular Differentiation System Maps Multiple Human Kidney Lineages from Pluripotent Stem Cells

Hiraku Tsujimoto,Tomoko Kasahara,Shin-Ichi Sueta,Toshikazu Araoka,Satoko Sakamoto,Chihiro Okada,Shin-Ichi Mae,Taiki Nakajima,Natsumi Okamoto,Daisuke Taura,Makoto Nasu,Tatsuya Shimizu,Makoto Ryosaka,Zhongwei Li,Masakatsu Sone,Makoto Ikeya,Akira Watanabe,Kenji Osafune

doi:10.1016/j.celrep.2020.03.040

Hiraku Tsujimoto, Tomoko Kasahara + Show 16 more

Open Access

https://doi.org/10.1016/j.celrep.2020.03.040

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Abstract

Recent studies using human pluripotent stem cells (hPSCs) have developed protocols to induce kidney-lineage cells and reconstruct kidney organoids. However, the separate generation of metanephric nephron progenitors (NPs), mesonephric NPs, and ureteric bud (UB) cells, which constitute embryonic kidneys, in invitro differentiation culture systems has not been fully investigated. Here, we create a culture system in which these mesoderm-like cell types and paraxial and lateral plate mesoderm-like cells are separately generated from hPSCs. We recapitulate nephrogenic niches from separately induced metanephric NP-like and UB-like cells, which are subsequently differentiated into glomeruli, renal tubules, and collecting ducts invitro and further vascularized invivo. Our selective differentiation protocols should contribute to understanding the mechanisms underlying human kidney development and disease and also supply cell sources for regenerative therapies.

Highlights

In amniotes, the mesonephros is the embryonic kidney, and the more complex metanephros acts as the adult kidney
The separate generation of metanephric nephron progenitors (NPs), mesonephric NPs, and ureteric bud (UB) cells in in vitro differentiation culture systems or the recapitulation of nephrogenesis-like physiological interactions between metanephric NPs and UB cells that occur in embryonic kidneys has not been fully investigated
To induce mesoderm from PSCs, we tested various concentrations of a glycogen synthase kinase 3b (GSK3b) inhibitor, CHIR99021, which is a key regulator of mesoderm induction, and monitored the expression of a primitive streak (PS) marker, BRACHYURY, on an human induced pluripotent stem cells (hiPSCs) line, 585A1 (Evans et al, 2012; Mendjan et al, 2014; Okita et al, 2013)

Summary

Introduction

The mesonephros is the embryonic kidney, and the more complex metanephros acts as the adult kidney. The mammalian metanephros develops by mutual interactions between two progenitor tissues, the ureteric bud (UB) and the metanephric mesenchyme (MM) (Costantini and Kopan, 2010; Taguchi et al, 2014). Several groups including ours have reported induction protocols for kidney progenitor cells from human pluripotent stem cells (hPSCs) (Mae et al, 2018; Morizane et al, 2015; Taguchi et al, 2014; Taguchi and Nishinakamura, 2017; Takasato et al, 2014, 2015; Toyohara et al, 2015; Xia et al, 2013). Taguchi et al (2014) found that inducing axial stem cells and the posterior intermediate mesoderm (IM) could facilitate the derivation of metanephric nephron progenitors (NPs) by using an embryoid body culture system.

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