Journal
STEM CELL REPORTS
Volume 2, Issue 6, Pages 751-760Publisher
CELL PRESS
DOI: 10.1016/j.stemcr.2014.04.016
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Funding
- Center for Medical System Innovation
- Graduate Program for Leaders in Life Innovation
- Core-to-Core Program A (Advanced Research Networks)
- Funding Program for World-Leading Innovative R&D on Science and Technology
- Center for NanoBio Integration
- S-innovation program, Nakayama Foundation for Human Science
- Nakatomi Foundation Research Grant
- [23689079]
- Grants-in-Aid for Scientific Research [14J01692] Funding Source: KAKEN
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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-(4methoxyphenyl) pyrido[40,30: 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.
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