Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction.

Abstract

Bone healing is a complex and well-organized process, regulated by various factors ranging from growth factors to hormones, cytokines, mechanical stimuli, and aging. Recently, numerous techniques have been devised to efficiently induce the differentiation of human induced pluripotent stem cells (hiPSCs) to osteoblasts. However, enhancing the efficiency of osteoblast differentiation remains a challenge. Thus, we induced the differentiation of hiPSCs to mesodermal cells through Wnt/BMP signaling based on the generation of hiPSCs. After successful generation of hiPSCs, we induced the differentiation of mesodermal cells to osteoblasts. The results revealed that the runt-related transcription factor 2 (RUNX2)-encoding gene was upregulated from the early differentiation stage; hence, the expression of the mature osteoblast marker was higher compared to that observed in other differentiation stages. In addition, the deposition of substrates in mature bones was observed. The results were confirmed via real-time PCR, Alizarin Red Staining, and Von Kossa staining. The coactivation of Wnt and BMP signaling was shown to rapidly and effectively promote the differentiation of osteoblasts. The findings of this study will provide a foundation for future studies on the mechanism of osteoblast development, as well as the biological and pathological investigations of drug screening and bone regeneration tracing.