[PDF][PDF] Interleukin-1 beta-modulated gene expression in immortalized human chondrocytes.

MB Goldring, JR Birkhead, LF Suen… - The Journal of …, 1994 - Am Soc Clin Investig
MB Goldring, JR Birkhead, LF Suen, R Yamin, S Mizuno, J Glowacki, JL Arbiser, JF Apperley
The Journal of clinical investigation, 1994Am Soc Clin Investig
Immortalized human chondrocytes were established by transfection of primary cultures of
juvenile costal chondrocytes with vectors encoding simian virus 40 large T antigen and
selection in suspension culture over agarose. Stable cell lines were generated that exhibited
chondrocyte morphology, continuous proliferative capacity (> 80 passages) in monolayer
culture in serum-containing medium, and expression of mRNAs encoding chondrocyte-
specific collagens II, IX, and XI and proteoglycans in an insulin-containing serum substitute …
Immortalized human chondrocytes were established by transfection of primary cultures of juvenile costal chondrocytes with vectors encoding simian virus 40 large T antigen and selection in suspension culture over agarose. Stable cell lines were generated that exhibited chondrocyte morphology, continuous proliferative capacity (> 80 passages) in monolayer culture in serum-containing medium, and expression of mRNAs encoding chondrocyte-specific collagens II, IX, and XI and proteoglycans in an insulin-containing serum substitute. They did not express type X collagen or versican mRNA. These cells synthesized and secreted extracellular matrix molecules that were reactive with monoclonal antibodies against type II collagen, large proteoglycan (PG-H, aggrecan), and chondroitin-4- and chondroitin-6-sulfate. Interleukin-1 beta (IL-1 beta) decreased the levels of type II collagen mRNA and increased the levels of mRNAs for collagenase, stromelysin, and immediate early genes (egr-1, c-fos, c-jun, and jun-B). These cell lines also expressed reporter gene constructs containing regulatory sequences (-577/+3,428 bp) of the type II collagen gene (COL2A1) in transient transfection experiments, and IL-1 beta suppressed this expression by 50-80%. These results show that immortalized human chondrocytes displaying cartilage-specific modulation by IL-1 beta can be used as a model for studying normal and pathological repair mechanisms.
Images
The Journal of Clinical Investigation