Stepwise Differentiation of Pluripotent Stem Cells into Osteoblasts Using Four Small Molecules under Serum-free and Feeder-free Conditions

• DOI: 10.1016/j.stemcr.2014.04.016
• Publication Date: 2014-05-22T16:22:22Z
• Authors: Kosuke Kanke; Hideki Masaki; Taku Saito; Yuske Komiyama; Hironori Hojo; Hiromitsu Nakauchi; Alexander C. Lich...; Tsuyoshi Takato; Ung-il Chung; Shinsuke Ohba
• Keywords: RUNX2

Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies.Here, we report an effective mass-producing strategy the stepwise differentiation mouse embryonic (mESCs) human induced pluripotent (miPSCs hiPSCs, respectively) into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], helioxanthin-derivative 4-(4-methoxyphenyl)pyrido[4 0 ,3 :4,5]thieno [2,3-b]pyridine-2-carboxamide [TH]) under serum-free feeder-free conditions.The strategy, which consists mesoderm induction, osteoblast maturation phases, significantly expressions osteoblast-related genes proteins in mESCs, miPSCs, hiPSCs.In addition, when mESCs defective runt-related transcription factor 2 (Runx2), master regulator osteogenesis, were cultured by they molecularly recapitulated phenotypes Runx2 null mice.The present will be platform biological pathological screening bone-augmentation drugs, skeletal regeneration.