Efficient and rapid induction of human iPSCs/ESCs into nephrogenic intermediate mesoderm using small molecule-based differentiation methods

PLoS One. 2014 Jan 15;9(1):e84881. doi: 10.1371/journal.pone.0084881. eCollection 2014.

Abstract

The first step in developing regenerative medicine approaches to treat renal diseases using pluripotent stem cells must be the generation of intermediate mesoderm (IM), an embryonic germ layer that gives rise to kidneys. In order to achieve this goal, establishing an efficient, stable and low-cost method for differentiating IM cells using small molecules is required. In this study, we identified two retinoids, AM580 and TTNPB, as potent IM inducers by high-throughput chemical screening, and established rapid (five days) and efficient (80% induction rate) IM differentiation from human iPSCs using only two small molecules: a Wnt pathway activator, CHIR99021, combined with either AM580 or TTNPB. The resulting human IM cells showed the ability to differentiate into multiple cell types that constitute adult kidneys, and to form renal tubule-like structures. These small molecule differentiation methods can bypass the mesendoderm step, directly inducing IM cells by activating Wnt, retinoic acid (RA), and bone morphogenetic protein (BMP) pathways. Such methods are powerful tools for studying kidney development and may potentially provide cell sources to generate renal lineage cells for regenerative therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation*
  • Cell Line
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Flow Cytometry
  • High-Throughput Screening Assays
  • Humans
  • Kidney / cytology*
  • Kidney / metabolism
  • Mesoderm / cytology*
  • Mice
  • Mice, Inbred ICR
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Signal Transduction

Grant support

This study was supported in part by research grants from the Leading Project of MEXT, the Uehara Memorial Foundation and the Takeda Science Foundation, by the Japan Society for the Promotion of Science (JSPS) through its "Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST Program)" and Grant-in-Aid for Young Scientists (B) to KO, and by Japan Science and Technology Agency (JST) through PRESTO to KO, JST Yamanaka iPS Cell Special Project to AO, SY and KO, and its research grant "Projects for Technological Development, Research Center Network for Realization of Regenerative Medicine". SM was supported by a fellowship from JSPS. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.