Expression profile of protein tyrosine kinase genes in human osteoarthritis chondrocytes

Abstract

Our results provide novel information about protein kinase gene expression in OA chondrocytes and indicate that the transformed chondrocyte cell line T/C 28a4 may be suitable for elucidating signal transduction pathways in chondrocytes and to investigate how they regulate chondrocyte function in inflammatory and degenerative joint diseases.

Phenotypic Characterization of Human Chondrocyte Cell Line C-20/A4: A Comparison between Monolayer and Alginate Suspension Culture

“…The C-20/A4 cells, which are neomycin-sensitive, have been particularly useful for stable expression of a gene of interest [Labelle et al, 1999;Attur et al, 2000;Nuttall et al, 2000;Laflamme et al, 2003]. Finally, immortalized chondrocyte cell lines have been used successfully to detect potential target genes of hydrostatic pressure and protein kinase profiles for further study by cDNA array analysis [Sironen et al, 2000;Islam et al, 2001]. Thus, immortalized chondrocyte cell lines, including the C-20/A4 cells, when cultured under the appropriate conditions have the potential in a variety of ways to extend the findings using primary cells or cartilage tissue cultures.…”

Interleukin-10 does not affect IL-1-induced interleukin-6 and metalloproteinase production in human chondrosarcoma cells, SW1353

“…The cartilage used for primary cultures of healthy tissue was obtained from the joints of legs amputated because of trauma or vascular disease after obtaining approval from the local ethics committee. Chondrocytes were prepared from macroscopically normal cartilage samples by enzymatic digestion, essentially as previously described (15). Isolated chondrocytes were plated in 75-cm 2 tissue culture flasks in Ham's F-12/DMEM (1:1) supplemented with 10% FCS, MEM vitamins, 100 μg/ml streptomycin, and 100 U/ml penicillin and cultured at 37˚C in humidified air with 10% CO 2 for a maximum of two passages.…”

Phenyl N-tert-Butylnitrone Down-Regulates Interleukin-1β-Stimulated Matrix Metalloproteinase-13 Gene Expression in Human Chondrocytes: Suppression of c-Jun NH2-Terminal Kinase, p38-Mitogen-Activated Protein Kinase and Activating Protein-1

“…Human OA cartilage samples from the hip were procured through the Cooperative Human Tissue Network and with prior approval of the Institutional Review Board of University Hospitals of Cleveland. In all of the experiments described in this article, primary chondrocytes prepared by the enzymatic digestion of cartilage (Singh et al, 2002;Islam et al, 2001) were used. OA chondrocytes were plated (1 ϫ 10 6 /ml) in 35-mm culture dishes (BD Biosciences, Franklin Lakes, NJ) in Dulbecco's modified Eagle's medium/F-12 (1:1) with 10% fetal calf serum (Invitrogen) and allowed to adhere for 72 h at 37°C and 5% CO 2 in a tissue culture incubator.…”