The Use of Laser Capture Microdissection on Adult Human Articular Cartilage for Gene Expression Analysis
The integrity of articular cartilage is maintained by chondrocytes, the sole type of cell that resides within the tissue. The noncalcified region of articular cartilage can be divided into three zones based on histological features, in which the chondrocyte metabolism is known to differ obviously among the zones. In pathological cartilage, the chondrocyte metabolism may change dramatically, which could play a pivotal role in the progression of the disease. Since such change in metabolism differs obviously from site to site within cartilage, it is crucial to determine the chondrocyte metabolism in respective regions. To this end, we have employed laser-capture microdissection (LCM) to analyze chondrocyte metabolism in various regions of pathological and control cartilage. In this report, we describe our protocol for LCM on adult human cartilage tissue. With this protocol, a specific site of cartilage tissue was successfully obtained by LCM for gene expression analysis.
Key wordsLaser-capture microdissection Articular cartilage mRNA Osteoarthritis Chondrocyte Metabolic change Regional difference
This work was sponsored in part by Grants-in-Aids from the Japan Society for the Promotion of Science (Nos. 15390467 and 18390424).
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