Three-dimensional perfusion culture of human bone marrow cells and generation of osteoinductive grafts

Stem Cells. 2005 Sep;23(8):1066-72. doi: 10.1634/stemcells.2005-0002. Epub 2005 Jul 7.


Three-dimensional (3D) culture systems are critical to investigate cell physiology and to engineer tissue grafts. In this study, we describe a simple yet innovative bioreactor-based approach to seed, expand, and differentiate bone marrow stromal cells (BMSCs) directly in a 3D environment, bypassing the conventional process of monolayer (two-dimensional [2D]) expansion. The system, based on the perfusion of bone marrow-nucleated cells through porous 3D scaffolds, supported the formation of stromal-like tissues, where BMSCs could be cocultured with hematopoietic progenitor cells in proportions dependent on the specific medium supplements. The resulting engineered constructs, when implanted ectopically in nude mice, generated bone tissue more reproducibly, uniformly, and extensively than scaffolds loaded with 2D-expanded BMSCs. The developed system may thus be used as a 3D in vitro model of bone marrow to study interactions between BMSCs and hematopoietic cells as well as to streamline manufacture of osteoinductive grafts in the context of regenerative medicine.

MeSH terms

  • Adult
  • Animals
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / ultrastructure
  • Cell Culture Techniques*
  • Cell Proliferation
  • Cells, Cultured
  • Culture Media / chemistry
  • Durapatite
  • Hematopoietic Stem Cells / cytology
  • Humans
  • Mesenchymal Stem Cells / cytology
  • Mice
  • Mice, Nude
  • Middle Aged
  • Osteogenesis*
  • Perfusion
  • RNA, Messenger / metabolism


  • Culture Media
  • RNA, Messenger
  • Durapatite