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Efficient and rapid induction of human iPSCs/ESCs into nephrogenic intermediate mesoderm using small molecule-based
Toshikazu Araoka1, Shin-ichi Mae1, Yuko Kurose1
1Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Researchers developed a rapid, efficient method to generate kidney precursor cells (intermediate mesoderm) from human stem cells using two small molecules. This breakthrough advances regenerative medicine for kidney disease treatment and research.
Area of Science:
- Stem cell biology
- Developmental biology
- Regenerative medicine
Background:
- Kidney disease treatment requires generating intermediate mesoderm (IM) from pluripotent stem cells.
- Current methods for IM differentiation are inefficient, unstable, or costly.
- Small molecule-based differentiation is needed for scalable IM generation.
Purpose of the Study:
- To establish an efficient, stable, and low-cost method for differentiating intermediate mesoderm (IM) cells from human induced pluripotent stem cells (iPSCs) using small molecules.
- To identify potent small molecules capable of inducing IM differentiation.
- To assess the potential of generated IM cells for kidney development and regenerative therapy.
Main Methods:
- High-throughput chemical screening to identify intermediate mesoderm (IM) inducers.
- Differentiation of human induced pluripotent stem cells (iPSCs) using a combination of CHIR99021 (Wnt activator) and retinoids (AM580 or TTNPB).
- Assessment of IM cell differentiation potential into kidney cell types and renal tubule-like structures.
Main Results:
- Identified AM580 and TTNPB as potent IM inducers.
- Established a rapid (5-day) and efficient (80% induction rate) IM differentiation protocol from human iPSCs.
- Demonstrated that generated human IM cells can differentiate into multiple kidney cell types and form renal tubule-like structures.
- Showcased that the method bypasses the mesendoderm stage by activating Wnt, retinoic acid (RA), and bone morphogenetic protein (BMP) pathways.
Conclusions:
- Developed a novel, efficient, and cost-effective small molecule-based method for inducing intermediate mesoderm (IM) from human iPSCs.
- The generated IM cells possess the potential for kidney development and can be utilized for regenerative therapies.
- This method offers a powerful tool for studying kidney development and generating renal lineage cells.
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