By Massimo Sabatini, Philippe Pastoureau, FrTdTric De Ceuninck
An vital choice of up to date classical and rising concepts that promise so as to add serious wisdom to our realizing of cartilage metabolism in well-being and disorder. quantity 1: mobile and Molecular instruments describes confirmed molecular and mobile recommendations for the in vitro examine of standard and osteoarthritic cartilage via biochemical, biomolecular, immunological, and actual techniques, with emphasis at the genetic manipulation of cells. quantity 2: constitution and In Vivo research, deals state-of-the-art systems for studies-at the tissue level-of turnover, constitution, and functioning of ordinary and diseased cartilage through invasive and noninvasive ability. accomplished and updated, the 2 volumes of Cartilage and Osteoarthritis supply researchers and bench scientists with without difficulty reproducible protocols for brand new experiments to understand-from the mobile to the animal level-the pathophysiology of cartilage and to find molecular goals for pharmacological intervention.
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Additional resources for Cartilage and Osteoarthritis
Total RNA was isolated and analyzed by RT-PCR for expression of COL2A1, aggrecan, and GAPDH mRNAs. Lane 1, DNA ladder; lane 2, primary adult articular chondrocytes; lane 3, T/C-28a4 cultured in medium with 10% FCS; lane 4, T/C28a4 in serum-free medium with 1% Nutridoma-SP; lane 5, C-28/I2 in 1% Nutridoma-SP; lane 6, C-28/I2 in 10% FCS; lane 7, C-20/A4 in 10% FCS; lane 8, tsT/AC62 in 10% FCS at 32°C. Note that some cell lines are susceptible to a loss of the COL2A1 phenotype in serumcontaining growth medium.
Biol. Chem. 272, 11,736–11,743. Chondrocyte Cell Lines As Culture Models 37 4 Culture of Immortalized Chondrocytes and Their Use As Models of Chondrocyte Function Mary B. Goldring Summary Immortalization of chondrocytes increases life span and proliferative capacity but does not necessarily stabilize the differentiated phenotype. Expansion of chondrocyte cell lines in continuous monolayer culture may result in the loss of phenotype, particularly if high cell density is not maintained. This chapter describes strategies for maintaining or restoring differentiated phenotype in established chondrocyte cell lines involving culture in serum-free defined culture medium, in suspension over agarose or polyHEMA, or within alginate or collagen scaffolds.
1. 30 Goldring Fig. 2. Representative growth curves from established immortalized chondrocyte cell lines expressing different forms of SV40-TAg. 5 × 104 cells/cm2 and cultured in DMEM/F-12 supplemented with 10% FCS for the indicated time periods with medium changes every 3 d. The T/C28a4, C-28/I2, and C-20/A4 cell lines are grown at 37°C. The tsT/AC62 cell line is grown at 32°C, the permissive temperature for expression of tsTAg. (Reproduced with permission from ref. ) 2. Growth kinetics: trypsinize the cells from a 10-cm dish and wash with PBS.
Cartilage and Osteoarthritis by Massimo Sabatini, Philippe Pastoureau, FrTdTric De Ceuninck