Senior Lecturer Okayama University Hospital Okayama, Okayama, Japan
Abstract: Human pluripotent stem cells (hPSCs) have shown great potential in recapitulating developmental processes for tissue engineering and disease modeling. However, generating functional skeletal tissues, especially those involved in limb development, remains a major challenge. In this study, we describe a novel methodology for differentiating human induced pluripotent stem cells (iPSCs) into region-specific skeletal progenitor cells, including those giving rise to articular cartilage and growth plate cartilage. We successfully induced limb bud mesenchymal cells (LBM) from iPSCs, derived from the lateral plate mesoderm, and used these cells as a foundation for generating articular cartilage and growth plate cartilage, two key structures in limb development. Through a combination of molecular signaling manipulations, 3D chondrogenic induction, and in vivo transplantation, we identified conditions that promote GDF5-positive (GDF5+) and GDF5-negative chondrogenic fates in a controlled manner. Moreover, we constructed human skeletal assembloids, combining chondrocyte progenitors from different regions of the limb, which mimicked endochondral ossification and longitudinal growth when transplanted into animal models. This study provides a significant advancement in stem cell-based limb regeneration and offers a powerful platform for investigating developmental biology and disease.
Funding Source: the JSPS(T.T and T.Takao) AMED (T.Takao) the JST FOREST Program (T.T) the Astellas Foundation for Research on Metabolic Disorders the Cooperative Research Program of the LiMe Kyoto University.
