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First published online 3 June 2003
doi: 10.1242/jcs.00527

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

¹ Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy
² Dipartimento di Medicina Sperimentale, Universita' dell'Aquila, Italy
³ Dipartimento di Medicina Sperimentale e Patologia, Universita' La Sapienza, Roma, Italy
⁴ Dipartimento di Oncologia, Biologia e Genetica, Universita' di Genova, Italy
⁵ Parco Scientifico Biomedico San Raffaele Roma, Italy

^* Author for correspondence (e-mail: rodolfo.quarto{at}istge.it)

Accepted 1 April 2003

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 ß-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

Key words: Osteoprogenitors, Osteogenesis, Chondrogenesis, Bone-marrow stromal cells, Condensation

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