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Sustained In Vitro Expansion of Bone Progenitors Is Cell Density Dependent

^a Department of Chemical Engineering and Applied Chemistry;
^b Institute of Biomaterials and Biomedical Engineering;
^c Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, Canada

Key Words. Colony forming unit-osteoblast • Expansion • Osteoprogenitor • Dexamethasone • Self-renewal

Peter W. Zandstra, Ph.D., Institute of Biomaterials and Biomedical Engineering, Room 407, Roseburgh Building, 4 Taddle Creek Road, Toronto, Ontario, M5S 3G9, Canada. Telephone: 416-978-8888; Fax: 416-978-4317; e-mail: peter.zandstra{at}utoronto.ca

Osteogenic cells are an integral part of the dynamic tissue-remodeling process in bone and are potential tools for tissue engineering and cell-based therapies. We examined the role of glucocorticoids and cell density in the expansion of primary rat calvaria cell populations and osteoprogenitor subpopulations in adherent cell culture. Osteoprogenitor response to dexamethasone (dex, a synthetic glucocorticoid known to stimulate bone formation in vitro) supplementation and long-term osteoprogenitor cell proliferation and differentiation were quantified using functional (colony forming unit-osteoblast [CFU-O]) and phenotypic analyses. Although osteoprogenitor self-renewal occurred at both standard and high initiating cell densities, progenitor cell expansion (measured by changes in CFU-O number relative to input) was sustained and dramatically increased at high initiating cell densities (30-fold CFU-O expansion for standard-density cultures compared with a greater than 10,000-fold CFU-O expansion in high-density cultures). Cell density was also found to impact upon the potential of dex to recruit additional progenitors towards bone development. These multifaceted effects appeared to be independent of cell proliferation rates or population phenotypic expression. Together, our results emphasize a roll for cell-cell interactions and/or community effects in the control and maintenance of progenitor cells during in vitro culture.

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