Formation of a chondro-osseous rudiment in micromass cultures of human bone-marrow stromal cells

J Cell Sci. 2003 Jul 15;116(Pt 14):2949-55. doi: 10.1242/jcs.00527. Epub 2003 Jun 3.

Abstract

Bone-marrow stromal cells can differentiate into multiple mesenchymal lineages including cartilage and bone. When these cells are seeded in high-density 'pellet culture', they undergo chondrogenesis and form a tissue that is morphologically and biochemically defined as cartilage. Here, we show that dual chondro-osteogenic differentiation can be obtained in the same micromass culture of human bone-marrow stromal cells. Human bone-marrow stromal cells were pellet cultured for 4 weeks in chondro-inductive medium. Cartilage 'beads' resulting from the micromass culture were then subcultured for further 1-3 weeks in osteo-inductive medium. This resulted in the formation of a distinct mineralized bony collar around hyaline cartilage. During the chondrogenesis phase, type I collagen and bone sialoprotein were produced in the outer portion of the cartilage bead, which, upon subsequent exposure to beta-glycerophosphate, mineralized and accumulated extracellular bone sialoprotein and osteocalcin. Our modification of the pellet culture system results in the formation of a chondro-osseous 'organoid' structurally reminiscent of pre-invasion endochondral rudiments, in which a bony collar forms around hyaline cartilage. The transition from a cell culture to an organ culture dimension featured by our system provides a suitable model for the dissection of molecular determinants of endochondral bone formation, which unfolds in a precisely defined spatial and temporal frame

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Bone Marrow / metabolism
  • Bone Marrow Cells / cytology*
  • Cartilage / cytology
  • Cell Differentiation
  • Cells, Cultured
  • Child
  • Child, Preschool
  • Chondrogenesis
  • Collagen Type I / metabolism
  • Glycerophosphates / pharmacology
  • Humans
  • Immunohistochemistry
  • Infant
  • Microscopy, Electron
  • Middle Aged
  • Osteocalcin / metabolism
  • Osteogenesis
  • Sialoglycoproteins / metabolism
  • Stromal Cells / cytology*
  • Time Factors

Substances

  • Collagen Type I
  • Glycerophosphates
  • Sialoglycoproteins
  • Osteocalcin
  • beta-glycerophosphoric acid