Novel strategy for enrichment and isolation of osteoprogenitor cells from iPS cells and the effectiveness in vivo

2014 Fiscal Year Final Research Report

• Project/Area Number: 25861761
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional basic dentistry
• Research Institution: Tokyo Dental College
• Principal Investigator: SHINO HIROMI 東京歯科大学, 歯学部, 助教 (00445446)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: iPS細胞 / 骨芽細胞 / 骨細胞 / 骨芽細胞分化

Outline of Final Research Achievements

In this study, we developed a new method to stimulate osteogenic differentiation in tissue-nonspecific alkaline phosphatase (TNAP)-positive cells liberated from human induced pluripotent stem cells (hiPSCs)-derived embryoid bodies (EBs) with 14 days long. TNAP is a marker protein of osteolineage cells. We analyzed and isolated TNAP-positive by fluorescence-activated cell sorting. Treating hiPSCs-derived cells with a combination of FGF-2, IGF-1, and TGF-β generated TNAP-positive cells at high frequency. The isolated cells expressed high levels of osterix, which is an exclusive osteogenic marker. Culturing these TNAP-positive cells in osteoblast differentiation medium (OBM) led to the expression of various osteogenic markers. Furthermore, in OBM they were capable of generating many mineralized nodules with strong expression of receptor activator of NF-kappaB ligand and sclerostin (SOST). Real-time RT-PCR showed a significant increase in the expression of osteocyte marker genes.

Free Research Field

生化学