A Novel Strategy for Enrichment and Isolation of Osteoprogenitor Cells from Induced Pluripotent Stem Cells Based on Surface Marker Combination

摘要: 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 (hiPSCs)-derived embryoid bodies (EBs) with 14 days long TGF-β/IGF-1/FGF-2 treatment. TNAP is marker protein of osteolineage cells. We analyzed and isolated TNAP-positive E-cadherin-negative nonepithelial by fluorescence-activated cell sorting. Treating the combination transforming growth factor (TGF)-β, insulin-like (IGF)-1, fibroblast (FGF)-2 for greatly enhanced expression maximized frequency up 77.3%. The expressed high levels osterix, which an exclusive marker. Culturing these osteoblast medium (OBM) led runt-related transcription 2, type I collagen, bone sialoprotein, osteocalcin (OCN). These responded treatment activated vitamin D3 upregulating OCN. Furthermore, OBM they were capable generating many mineralized nodules strong receptor activator NF-kappaB ligand sclerostin (SOST). Real-time RT-PCR showed significant increase osteocyte genes, including SOST, neuropeptide Y, reelin. Scanning electron microscopy dendritic morphology. Examination semi-thin toluidine blue-stained sections interconnected dendrites. Thus, cultured may eventually become terminally differentiated osteocyte-like conclusion, treating hiPSCs-derived TGF-β, IGF-1, FGF-2 generated at frequency. had potential could differentiate into described here reveal pathways osteogenesis provide novel tool regenerative medicine drug development.