Making a DNA Chip for the Study of Cartilage, and a New Challenge for the Repair and Regeneration of Cartilage Tissue.

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In this research project, gene trap and cDNA subtraction methods were used to identify comprehensively the genes specifically expressed in cartilage. Furthermore, commercially available cDNA chip was also used for the analysis. Although tissue specific gene trap system that we have been originally developing for this project has not been completed yet, signal sequence specific gene trap vector, a byproduct of the system construction, has been working well to identify cell surface and extracellular matrix molecules produced by chondrocytes. Similarly, we found that cDNA subtraction analysis works well by selecting tissues or cells properly for a comparison of gene expression. For example, we compared the gene expression of articular cartilage and growth plate cartilage by suppression subtractive hybridization, and could identify several genes specifically expressed in articular cartilage but not in growth plate cartilage. One such molecule is lubricin that is specifically expressed on the superficial layer of articular cartilage, and has an important function for the lubrication of joint. Of the other molecules, we could confirm their specific expression in each cartilage by Northern blot analysis. However, cDNA microarray analysis using commercially available one did not give us fruitful results as we expected. This is due to the fact that commercially available cDNA chips are still imperfect for analyzing genes expressed in cartilage. Therefore, comprehensive analysis of genes expressed in cartilage is still very important, and the production of cartilage specific cDNA microarray is most urgent challenge for us.