Cryopreservation and in vitro expansion of chondroprogenitor cells isolated from the superficial zone of articular cartilage

Juan Melero Martin, M Smith, Mohamed Al-Rubeai

Research output: Contribution to journalArticle

20 Citations (Scopus)


Understanding the proliferation mechanisms of chondroprogenitor cells and their influence on cell differentiation is crucial in order to develop large-scale expansion processes for tissue engineering applications. Proliferation control mechanisms were mainly attributed to substrate limitation and cell-cell contact inhibition. The limiting substrates were found to be components of the FCS, with an optimal proliferation rate achieved in the presence of 40% FCS. In addition, the medium supply rate was found to be essential in reducing substrate limitation. In terms of FCS, 10 muL FCS cm(-2) h(-1) was the threshold feed rate required to prevent substrate limitation. Above this rate, maximum cell densities of 5.3 x 10(5) cells/cm(2) were achieved, representing a 53-fold expansion. To reduce the need for high supply rates, the effect of specific growth factors was also investigated. Cell densities of 3.3 x 10(5) cells/cm(2) were achieved in batch cultures using 40% FCS and 1 ng/mL TGF-beta1 Chondroprogenitor cells were expanded in this medium up to three passages without compromising their ability to differentiate and produce cartilage-like matrix in pellet cultures. In addition to substrate limitation, cell-cell contact, even at very sparse subconfluent densities, appeared capable of exerting some degree of growth inhibition. The cells exhibited deceleratory growth kinetics, characterized by a decrease of specific growth rates over time.
Original languageEnglish
Pages (from-to)168-177
Number of pages10
JournalBiotechnology Progress
Publication statusPublished - 1 Jan 2005


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