Cultivation of Human Bone-Like Tissue from Pluripotent Stem Cell-Derived Osteogenic Progenitors in Perfusion Bioreactors

  • Giuseppe Maria de Peppo
  • Gordana Vunjak-Novakovic
  • Darja MaroltEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1202)


Human pluripotent stem cells represent an unlimited source of skeletal tissue progenitors for studies of bone biology, pathogenesis, and the development of new approaches for bone reconstruction and therapies. In order to construct in vitro models of bone tissue development and to grow functional, clinical-size bone substitutes for transplantation, cell cultivation in three-dimensional environments composed of porous osteoconductive scaffolds and dynamic culture systems—bioreactors—has been studied. Here, we describe a stepwise procedure for the induction of human embryonic and induced pluripotent stem cells (collectively termed PSCs) into mesenchymal-like progenitors, and their subsequent cultivation on decellularized bovine bone scaffolds in perfusion bioreactors, to support the development of viable, stable bone-like tissue in defined geometries.


Human embryonic stem cells Human induced pluripotent stem cells Mesenchymal-like progenitors Osteogenic differentiation Osteogenic medium Bone scaffolds Cell seeding Perfusion bioreactor Medium flow rate Bone tissue development 



This work was supported by the New York Stem Cell Foundation-Helmsley Investigator Award (to D.M.); the Leona M. and Harry B. Helmsley Charitable Trust; Robin Chemers Neustein; Goldman Sachs Gives, at the recommendation of Alan and Deborah Cohen; New York State Stem Cell Science Shared Facility, Grant C024179; National Institutes of Health Grants DE016525 and EB002520, (to G.V.-N.); and the New York Stem Cell Foundation.


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Giuseppe Maria de Peppo
    • 2
  • Gordana Vunjak-Novakovic
    • 3
  • Darja Marolt
    • 1
    Email author
  1. 1.The New York Stem Cell Foundation Research InstituteNew YorkUSA
  2. 2.The New York Stem Cell Foundation Research InstituteNew YorkUSA
  3. 3.Department of Biomedical Engineering and Department of MedicineColumbia UniversityNew YorkUSA

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