Influence of medium composition, static and stirred conditions on the proliferation of and matrix protein expression of bovine articular chondrocytes cultured in a 3-D collagen scaffold.
Interest in chemical and physical modifications of culture conditions and composition, as a way to improve engineered cartilage, has grown over the last decade. To address some of these aspects, articular bovine chondrocytes seeded in collagen sponges (2.3x10(6) cells/cm(3), whose growth and metabolism have been previously reported) were grown under static or stirred conditions (orbital shaker at 30 rpm), in either 10% FCS-supplemented or serum-free media (1% ITS+1mM cysteine). Under stirred conditions, we observed a 2-fold increase in both cell proliferation and sulphated glycosaminoglycan deposition after 1 month of culture, compared to static conditions, and after 3 months, a more homogeneous distribution of both cells and neomatrix in the constructs. During the first month of culture, the substitution of FCS by ITS led to low cell proliferation and poor neomatrix deposition but, after 2 months a steep increase was observed with ITS for these two parameters that reached, after 3 months the levels observed with FCS. Aggrecan was the more abundant component at both gene and protein levels, whereas the collagenous network formed was looser than with FCS. In conclusion, the use of these simple culture conditions should improve, in long-term culture, the quality of the cartilage construct.
Freyria
A-M
AM
Institut de Biologie et Chimie des Protéines, UMR 5086, CNRS-Université Lyon I & IFR, 128-Biosciences Lyon-Gerland, 7 Passage du Vercors, 69367 Lyon Cedex 7, France. am.freyria@ibcp.fr
Cortial
D
D
Ronzière
M-C
MC
Guerret
S
S
Herbage
D
D
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology* / Q000502:physiology*; D002417:Cattle; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology*; D003094:Collagen / Q000737:chemistry*; D003470:Culture Media / Q000378:metabolism*; D016895:Culture Media, Serum-Free / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D008422:Materials Testing; D009038:Motion; D023822:Tissue Engineering / Q000379:methods*
D003470:Culture Media; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; D003094:Collagen