Effects of in vitro preculture on in vivo development of human engineered cartilage in an ectopic model

Matteo Moretti, David Wendt, Sally C. Dickinson, Trevor J. Sims, Anthony P. Hollander, Danny J. Kelly, Patrick J. Prendergast, Michael Heberer, Ivan Martin

Research output: Contribution to journalArticle

Abstract

We investigated whether, and under which conditions (i.e., cell-seeding density, medium supplements), in vitro preculture enhances in vivo development of human engineered cartilage in an ectopic nude mouse model. Monolayer-expanded adult human articular chondrocytes (AHACs) were seeded into Hyalograft C disks at 1.3 × 107 cells/cm3 (low density) or 7.6 × 107 cells/cm3 (high density). Constructs were directly implanted subcutaneously in nude mice for up to 8 weeks or precultured for 2 weeks before implantation. Preculture medium contained either transforming growth factor-β1 (TGF-β1, 1 ng/mL), fibroblast growth factor-2, and platelet-derived growth factor (proliferating medium) or TGF-β1 (10 ng/mL) and insulin (differentiating medium). Both in vitro and after in vivo implantation, constructs derived by cell seeding at high versus low density and precultured in differentiating versus proliferating medium generated more cartilaginous tissues containing higher amounts of glycosaminoglycan and collagen type II and lower amounts of collagen type I, and with higher equilibrium moduli. As compared with direct implantation of freshly seeded scaffolds, preculture of AHAC-Hyalograft C constructs in differentiating medium, but not in proliferating medium, supported enhanced in vivo development of engineered cartilage. The effect of preculture was more pronounced when constructs were seeded at low density as compared with high density. This study indicates that preculture of human engineered cartilage in differentiating medium has the potential to provide grafts with higher equilibrium moduli and enhanced in vivo developmental capacity than freshly seeded scaffolds. These findings need to be validated in an orthotopic model system.

Original languageEnglish
Pages (from-to)1421-1428
Number of pages8
JournalTissue Engineering
Volume11
Issue number9-10
DOIs
Publication statusPublished - Sep 2005

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Biotechnology

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    Moretti, M., Wendt, D., Dickinson, S. C., Sims, T. J., Hollander, A. P., Kelly, D. J., Prendergast, P. J., Heberer, M., & Martin, I. (2005). Effects of in vitro preculture on in vivo development of human engineered cartilage in an ectopic model. Tissue Engineering, 11(9-10), 1421-1428. https://doi.org/10.1089/ten.2005.11.1421