The recapitulation of bone formation via the in vitro generation of bone-like nodules is frequently used to understand bone development. However, current bone-induction techniques are slow and difficult to reproduce. Here, we report the formation of bone-like nodules within ten days, via the use of retinoic acid (RA) to induce the osteogenic differentiation of human induced pluripotent stem cells (hiPSCs) into osteoblast-like and osteocyte-like cells that create human bone tissue when implanted in calvarial defects in mice. We also show that the induction of bone formation depends on cell signalling through the RA receptors RARα and RARβ, which simultaneously activate the BMP (bone morphogenetic protein) and Wnt signalling pathways. Moreover, by using patient-derived hiPSCs, the bone-like nodules recapitulated the osteogenesis-imperfecta phenotype, which was rescued via the correction of disease-causing mutations and partially by an mTOR (mechanistic target of rapamycin) inhibitor. The method of inducing bone nodules may serve as a fast and reproducible model for the study of the formation of both healthy and pathological bone.
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The authors declare that all data supporting the results in this study are available within the paper and its Supplementary Information. Raw data are available from the corresponding author on reasonable request. The affymetrix data have been deposited in the GEO database, under accession number GSE119577.
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We thank Y. Matsumoto for his contribution in preliminary experiments; K. Hino for advice on the in vivo experiments and microarray analysis; M. Watanabe, K. Horigome and T. Takarada for invaluable comments and discussion; Y. Pretemer and R. Kashimoto for experimental supports; K. Woltjen for providing the 317–12 iPS cell line; A. Hotta and H. Xu for advice on gene editing; and P. Karagiannis for reading the manuscript. Preparation of the microscope slides was supported by staff at the Center for Anatomical, Pathological and Forensic Medical Research, Graduate School of Medicine, Kyoto University. This work was supported by Grants-in-aid for Scientific Research from Japan Society for the Promotion of Science (no. 17K19725), Research Center Network for Realization of Regenerative Medicine, The Core Center for iPS Cell Research and The Program for Intractable Diseases Research utilizing Disease-specific iPS from Japan Science and Technology Agency and Japan Agency for Medical Research and Development (AMED), The Acceleration Program for Intractable Diseases Research utilizing Disease-specific iPS cells from AMED to J.T., and The Advanced Research and Development Programs for Medical Innovation (CREST) from AMED to J.S. and T.A. These funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
The authors declare no competing interests.
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Kawai, S., Yoshitomi, H., Sunaga, J. et al. In vitro bone-like nodules generated from patient-derived iPSCs recapitulate pathological bone phenotypes. Nat Biomed Eng 3, 558–570 (2019). https://doi.org/10.1038/s41551-019-0410-7
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