The aim of this study was to establish the potential of human periosteum-derived cells from elderly patients as a cell source for cartilage tissue engineering by optimizing culture conditions for both proliferation and differentiation. Periosteum was obtained from the tibiae of nine patients. Biopsies were prepared for routine histological examination. Periosteum-derived cells were allowed to grow out from the remaining tissue, and were expanded in minimum essential medium containing D-valine (MEM-DV). Fetal bovine serum (FBS) or substitutes, fibroblast growth factor-2 (FGF-2), insulin-like growth factor-1 (IGF-1) and non-essential amino acids were added to study proliferation. For differentiation of cells, serum-free medium was used supplemented with one or more isoforms of transforming growth factor-beta (TGFbeta) and/or IGF-1. Samples were analysed for expression of collagens type I, II and X by competitive RT-PCR, immunohistochemically, and histologically using Alcian blue staining. In all samples the cambium layer could hardly be detected. Periosteum-derived cells proliferated in serum-containing MEM-DV. Optimal proliferation was found when this medium was supplemented with 100 ng/ml FGF-2 and non-essential amino acids. Chondrogenesis was detected in 59% of micromasses that were cultured with TGFbeta isomers, and in 83% of the samples cultured in media to which two TGFbeta isoforms were added. Periosteum from elderly humans (mean age 66, range 41-76 years) has chondrogenic potential and remains an attractive cell source for cartilage tissue engineering. By expanding cells in MEM-DV, the selection of progenitor cells might be favoured, which would result in a higher cartilage yield for tissue engineering applications.
(c) 2008 John Wiley & Sons, Ltd.