Isolation of RNA from small human articular cartilage specimens allows quantification of mRNA expression levels in local articular cartilage defects

• Published in: Journal of orthopaedic research Vol. 19; no. 3; pp. 478 - 481
• Main Authors: Gehrsitz, Angelika, McKenna, Louise A, Söder, Stefan, Kirchner, Thomas, Aigner, Thomas
• Format: Journal Article
• Language: English
• Published: Hoboken Elsevier Ltd 01.05.2001; Wiley Subscription Services, Inc., A Wiley Company; Blackwell Publishing Ltd
• Subjects: aggrecan; Aggrecans; Cartilage, Articular - chemistry; Cartilage, Articular - pathology; Collagen - genetics; Collagen - metabolism; DNA Primers - chemistry; Extracellular Matrix Proteins; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics; Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism; Humans; Knee Joint; Lectins, C-Type; Middle Aged; Polymerase Chain Reaction; Proteoglycans - genetics; Proteoglycans - metabolism; Reproducibility of Results; RNA - isolation & purification; RNA, Messenger - biosynthesis
• ISSN: 0736-0266; 1554-527X
• DOI: 10.1016/S0736-0266(00)90028-7

Human adult cartilage is an inherently difficult tissue from which to isolate RNA. The RNA isolation techniques described so far have generally only been successfully applied to the isolation of RNA from larger amounts of cartilage. However, it is important to be able to analyse focal cartilage lesions in order to understand the local processes in the cartilage degeneration process. Therefore, we have developed a protocol for isolating RNA directly from as little as 10 mg wet weight of cartilage followed by quantitative PCR analysis. We were able to analyse the expression levels of several genes in parallel including aggrecan and type II collagen.