title: Generation of Directly Converted Human Osteoblasts That Are Free of Exogenous Gene and Xenogenic Protein.
abstract: Generation of osteoblasts from human somatic cells may be applicable in an effective transplantation therapy against bone diseases. Recently we established a procedure to directly convert human fibroblasts into osteoblasts by transducing some transcription factor genes via retroviral vectors. However, retroviral vector-mediated transduction may potentially cause tumor formation from the infected cells, thus a non-viral gene transfection method may be more preferable for preparation of osteoblasts to be used for transplantation therapy. Here, we constructed a plasmid vector encoding Oct4, Osterix, and L-Myc that were an appropriate combination of transcription factors for this purpose. Osteoblast-like phenotypes including high alkaline phosphatase (ALP) activity, bone matrix production and osteoblast-specific gene expression were induced in normal human fibroblasts that were transfected with the plasmid followed by culturing in osteogenic medium. The plasmid-driven directly converted osteoblasts (p-dOBs) were obtained even in the absence of a xenogenic protein. The plasmid vector sequence had fallen out of the p-dOBs. The cells formed deposition of calcified bodies in situ after transplantation into mice. These results strongly suggest that p-dOBs can be put into practical use for a novel cell-based therapy against bone diseases. J. Cell. Biochem. 117: 2538-2545, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan.|Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan.|Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sato
authors.affiliation: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Honjo
authors.affiliation: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ichioka
authors.affiliation: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Fumishige
authors.initials: F
authors.lastname: Oseko
authors.affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan.|Department of Plastic and Reconstructive Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Sowa
authors.affiliation: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Toshiro
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Narisato
authors.initials: N
authors.lastname: Kanamura
authors.affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Tsunao
authors.initials: T
authors.lastname: Kishida
authors.affiliation: Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan. mazda@koto.kpu-m.ac.jp.
authors.forename: Osam
authors.initials: O
authors.lastname: Mazda
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism*; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: C000596782:MYCL protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D016271:Proto-Oncogene Proteins c-myc; D000076182:Sp7 Transcription Factor; C478163:SP7 protein, human; D014157:Transcription Factors
keywords: DIRECT REPROGRAMMING; OSTEOBLASTS; PLASMID VECTOR; REGENERATIVE MEDICINE