Osteoblasts Derived from Induced Pluripotent Stem Cells form Calcified Structures in Scaffolds Both In Vitro and In Vivo§

Authors

  • Ganna Bilousova,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Dermatology, University of Colorado Denver, Aurora, Colorado, USA
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  • Du Hyun Jun,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Medicine, Cardiovascular Pulmonary Research Laboratory, University of Colorado Denver, Aurora, Colorado, USA
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  • Karen B. King,

    1. Department of Orthopaedics, Division of Bioengineering, University of Colorado Denver, Aurora, Colorado, USA
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  • Stijn De Langhe,

    1. Department of Pediatrics, Division of Cell Biology, National Jewish Health, Denver, Colorado, USA
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  • Wallace S. Chick,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
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  • Enrique C. Torchia,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Dermatology, University of Colorado Denver, Aurora, Colorado, USA
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  • Kelsey S. Chow,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Medicine, Cardiovascular Pulmonary Research Laboratory, University of Colorado Denver, Aurora, Colorado, USA
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  • Dwight J. Klemm,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Medicine, Cardiovascular Pulmonary Research Laboratory, University of Colorado Denver, Aurora, Colorado, USA
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  • Dennis R. Roop,

    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Dermatology, University of Colorado Denver, Aurora, Colorado, USA
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  • Susan M. Majka

    Corresponding author
    1. Charles C. Gates Regenerative Medicine and Stem Cell Biology Program, University of Colorado Denver, Aurora, Colorado, USA
    2. Department of Medicine, Cardiovascular Pulmonary Research Laboratory, University of Colorado Denver, Aurora, Colorado, USA
    • University of Colorado Denver Health Sciences Center, Gates Center for Regenerative Medicine and Stem Cell Biology, 12,800 E 1nineth Ave, P.O. Box 6511, Mail stop 8320, Aurora, Colorado 80045, USA
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    • Telephone: 303-724-3957; Fax: 303-724-3051


  • G.B.: conception and design, collection and/or assembly of data, provision of study material or patients data analysis and interpretation, manuscript writing; D.H.J.: conception and design, collection and/or assembly of data, data analysis and interpretation; K.B.K.: provision of study material or patients, data analysis and interpretation, final approval of manuscript; S.D.L.: collection and/or assembly of data; W.S.C.: conception and design; E.C.T.: collection and/or assembly of data, data analysis and interpretation; K.S.C.: collection and/or assembly of data, data analysis and interpretation; D.J.K.: collection and/or assembly of data, data analysis and interpretation; D.R.R.: financial support, provision of study material or patients; S.M.M.: conception and design, financial support, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript. G.B. and D.H.J. contributed equally to this article.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLSEXPRESS November 23, 2010.

Abstract

Reprogramming somatic cells into an ESC-like state, or induced pluripotent stem (iPS) cells, has emerged as a promising new venue for customized cell therapies. In this study, we performed directed differentiation to assess the ability of murine iPS cells to differentiate into bone, cartilage, and fat in vitro and to maintain an osteoblast phenotype on a scaffold in vitro and in vivo. Embryoid bodies derived from murine iPS cells were cultured in differentiation medium for 8–12 weeks. Differentiation was assessed by lineage-specific morphology, gene expression, histological stain, and immunostaining to detect matrix deposition. After 12 weeks of expansion, iPS-derived osteoblasts were seeded in a gelfoam matrix followed by subcutaneous implantation in syngenic imprinting control region (ICR) mice. Implants were harvested at 12 weeks, histological analyses of cell and mineral and matrix content were performed. Differentiation of iPS cells into mesenchymal lineages of bone, cartilage, and fat was confirmed by morphology and expression of lineage-specific genes. Isolated implants of iPS cell-derived osteoblasts expressed matrices characteristic of bone, including osteocalcin and bone sialoprotein. Implants were also stained with alizarin red and von Kossa, demonstrating mineralization and persistence of an osteoblast phenotype. Recruitment of vasculature and microvascularization of the implant was also detected. Taken together, these data demonstrate functional osteoblast differentiation from iPS cells both in vitro and in vivo and reveal a source of cells, which merit evaluation for their potential uses in orthopedic medicine and understanding of molecular mechanisms of orthopedic disease. STEM CELLS 2011;29:206–216

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