Abstract
Implantation of chondrocytes isolated from patients and expanded in number in vitro is being used to treat patients with cartilage injuries. However, chondrocytes de-differentiate during culture with several passages, and cartilage regenerated by implantation of de-differentiated chondrocytes may be suboptimal. Here, we show that a spinner-flask culture system induces formation of chondrocyte aggregates and redifferentiate de-differentiated chondrocytes. Spinner-flask cultures induced the aggregate formation of chondrocytes with passage 1 or 4. Importantly, spinner-flask cultures induced redifferentiation of the de-differentiated chondrocytes, as type I collagen expression was significantly lower and type II collagen expression was significantly higher in spinner flask-cultured chondrocytes than in monolayer-cultured chondrocytes. This system is easily scalable and could be feasible for clinical setting.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bernstein P, Dong M, Corbeil D et al (2009) Pellet culture elicits superior chondrogenic redifferentiation than alginate-based systems. Biotechnol Prog 25:1146–1152
Capito RM, Spector M (2006) Effect of expansion medium on ex vivo gene transfer and chondrogenesis in type II collagen-glycosaminoglycan scaffolds in vitro. Osteoarthr Cartil 14:1203–1213
Duval E, Leclercq S, Elissalde JM et al (2009) Hypoxia-inducible factor 1alpha inhibits the fibroblast-like markers type I and type III collagen during hypoxia-induced chondrocyte redifferentiation: hypoxia not only induces type II collagen and aggrecan, but it also inhibits type I and type III collagen in the hypoxia-inducible factor 1alpha-dependent redifferentiation of chondrocytes. Arthritis Rheum 60:3038–3048
Heywood HK, Lee DA (2010) Low oxygen reduces the modulation to an oxidative phenotype in monolayer-expanded chondrocytes. J Cell Physiol 222:248–253
Kang SW, Yoon JR, Lee JS et al (2006) The use of poly(lactic-co-glycolic acid) microspheres as injectable cell carriers for cartilage regeneration in rabbit knees. J Biomater Sci Polym Ed 17:925–939
Kessler MW, Grande DA (2008) Tissue engineering and cartilage. Organogenesis 4:28–32
Lin Z, Fitzgerald JB, Xu J et al (2008) Gene expression profiles of human chondrocytes during passaged monolayer cultivation. J Orthop Res 26:1230–1237
Liu G, Kawaguchi H, Ogasawara T et al (2007) Optimal combination of soluble factors for tissue engineering of permanent cartilage from cultured human chondrocytes. J Biol Chem 282:20407–20415
Malda J, Kreijveld E, Temenoff JS et al (2003) Expansion of human nasal chondrocytes on macroporous microcarriers enhances redifferentiation. Biomaterials 24:5153–5161
Melero-Martin JM, Dowling MA, Smith M et al (2006) Expansion of chondroprogenitor cells on macroporous microcarriers as an alternative to conventional monolayer systems. Biomaterials 27:2970–2979
Ryu JH, Kim MS, Lee GM et al (2005) The enhancement of recombinant protein production by polymer nanospheres in cell suspension culture. Biomaterials 26:2173–2181
Schipani E, Ryan HE, Didrickson S et al (2001) Hypoxia in cartilage: HIF-1alpha is essential for chondrocyte growth arrest and survival. Genes Dev 15:2865–2876
Stevens MM, Qanadilo HF, Langer R et al (2004) A rapid-curing alginate gel system: utility in periosteum-derived cartilage tissue engineering. Biomaterials 25:887–894
Sung LY, Chiu HY, Chen HC et al (2009) Baculovirus-mediated growth factor expression in de-differentiated chondrocytes accelerates redifferentiation: effects of combinational transduction. Tissue Eng Part A 15:1353–1362
Takahashi T, Ogasawara T, Asawa Y et al (2007) Three-dimensional microenvironments retain chondrocyte phenotypes during proliferation culture. Tissue Eng 13:1583–1592
Veilleux NH, Yannas IV, Spector M (2004) Effect of passage number and collagen type on the proliferative, biosynthetic, and contractile activity of adult canine articular chondrocytes in type I and II collagen-glycosaminoglycan matrices in vitro. Tissue Eng 10:119–127
Villar-Suárez V, Calles-Venal I, Bravo IG et al (2004) Differential behavior between isolated and aggregated rabbit auricular chondrocytes on plastic surfaces. J Biomed Biotechnol 2004:86–92
Vinatier C, Gauthier O, Fatimi A et al (2009) An injectable cellulose-based hydrogel for the transfer of autologous nasal chondrocytes in articular cartilage defects. Biotechnol Bioeng 102:1259–1267
Yang KG, Saris DB, Geuze RE et al (2006) Impact of expansion and redifferentiation conditions on chondrogenic capacity of cultured chondrocytes. Tissue Eng 12:2435–2447
Acknowledgments
This study was supported by a grant (2009-0086518 and 2009-0092213) from the National Research Foundation of Korea, Republic of Korea.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, TJ., Bhang, S.H., La, WG. et al. Spinner-flask culture induces redifferentiation of de-differentiated chondrocytes. Biotechnol Lett 33, 829–836 (2011). https://doi.org/10.1007/s10529-010-0488-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-010-0488-1