title: Primary bone-derived cells induce osteogenic differentiation without exogenous factors in human embryonic stem cells.
abstract: We developed a new and efficient method for osteoblastic differentiation of human embryonic stem cells (hESCs) using primary bone-derived cells (PBDs). Three days after embryoid body (hEB) formation, cells were allowed to adhere to culture surface where PBDs were pre-plated and mitomycin C-treated in DMEM/F12 medium supplemented with 5% knockout serum replacement. As early as 14 days, mineralization and formation of nodule-like structures in cocultured hEBs were prominent by von Kossa and Alizarin S staining, and expressions of osteoblast-specific markers including bone sialoprotein, alkaline phosphates, osteocalcin, collagen 1, and core binding factor alpha1 by RT-PCR. In addition, FACS analysis revealed that over 19% of the differentiated cells expressed osteocalcin. These results suggest that PBDs not only have osteogenic effects releasing osteogenic factors as bone morphogenic protein (BMP) 2 and BMP 4 but also have exerted other effects, whether chemical or physical, for the differentiation of hESCs.
authors.affiliation: Stem Cell Research Laboratory, Cell and Gene Therapy Research Institute, Pochon CHA University, Seoul 135-081, Republic of Korea.
authors.forename: Seong Eun
authors.initials: SE
authors.lastname: Ahn
authors.forename: Sinae
authors.initials: S
authors.lastname: Kim
authors.forename: Kyu Hyung
authors.initials: KH
authors.lastname: Park
authors.forename: Sung Hwan
authors.initials: SH
authors.lastname: Moon
authors.forename: Hae Jin
authors.initials: HJ
authors.lastname: Lee
authors.forename: Gi Jin
authors.initials: GJ
authors.lastname: Kim
authors.forename: Young Jae
authors.initials: YJ
authors.lastname: Lee
authors.forename: Keun Hong
authors.initials: KH
authors.lastname: Park
authors.forename: Kwang Yul
authors.initials: KY
authors.lastname: Cha
authors.forename: Hyung Min
authors.initials: HM
authors.lastname: Chung
mesh_terms: D000328:Adult; D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis / Q000235:genetics; D001842:Bone and Bones / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D006133:Growth Substances; D006801:Humans; D010012:Osteogenesis / Q000235:genetics / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics
chemical_list: C525718:BMP2 protein, human; C525809:BMP4 protein, human; D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D006133:Growth Substances; D016212:Transforming Growth Factor beta