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Updated: Apr 28, 2026
Differentiation and Characterization of Osteoclasts from Human Induced Pluripotent Stem Cells
Published on: March 22, 2024
Stepwise differentiation of pluripotent stem cells into osteoblasts using four small molecules under serum-free and
Kosuke Kanke1, Hideki Masaki2, Taku Saito3
1Center for Disease Biology and Integrative Medicine, The University of Tokyo, Tokyo 113-0033, Japan ; Department of Sensory and Motor System Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
Abstract:
Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies. Here, we report an effective and mass-producing strategy for the stepwise differentiation of mouse embryonic stem cells (mESCs) and mouse and human induced pluripotent stem cells (miPSCs and hiPSCs, respectively) into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], and a helioxanthin-derivative 4-(4-methoxyphenyl)pyrido[4',3':4,5]thieno[2,3-b]pyridine-2-carboxamide [TH]) under serum-free and feeder-free conditions. The strategy, which consists of mesoderm induction, osteoblast induction, and osteoblast maturation phases, significantly induced expressions of osteoblast-related genes and proteins in mESCs, miPSCs, and hiPSCs. In addition, when mESCs defective in runt-related transcription factor 2 (Runx2), a master regulator of osteogenesis, were cultured by the strategy, they molecularly recapitulated osteoblast phenotypes of Runx2 null mice. The present strategy will be a platform for biological and pathological studies of osteoblast development, screening of bone-augmentation drugs, and skeletal regeneration.
