Efficient and rapid induction of human iPSCs/ESCs into nephrogenic intermediate mesoderm using small molecule-based differentiation methods.
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.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Toshikazu
T
Araoka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Shin-ichi
S
Mae
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Yuko
Y
Kurose
Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan ; Institute for Chemical Research, Kyoto University, Kyoto, Japan.
Motonari
M
Uesugi
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Akira
A
Ohta
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan ; Gladstone Institute of Cardiovascular Disease, San Francisco, California, United States of America.
Shinya
S
Yamanaka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Kenji
K
Osafune
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D057166:High-Throughput Screening Assays; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D008813:Mice, Inbred ICR; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D015398:Signal Transduction