在成人人类关节软骨上使用激光捕获显微切割技术进行基因表达分析。

The use of laser capture microdissection on adult human articular cartilage for gene expression analysis.

作者信息

Fukui Naoshi, Ikeda Yasuko, Tanaka Nobuho

机构信息

Clinical Research Center, National Hospital Organization, Sagamihara Hospital, Sagamihara, Kanagawa, Japan.

出版信息

Methods Mol Biol. 2011;755:449-59. doi: 10.1007/978-1-61779-163-5_38.

DOI:10.1007/978-1-61779-163-5_38

PMID:21761327

Abstract

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.

摘要

关节软骨的完整性由软骨细胞维持，软骨细胞是存在于该组织中的唯一细胞类型。根据组织学特征，关节软骨的非钙化区域可分为三个区，已知软骨细胞代谢在这些区域之间存在明显差异。在病理性软骨中，软骨细胞代谢可能会发生显著变化，这可能在疾病进展中起关键作用。由于软骨内不同部位的这种代谢变化明显不同，因此确定各个区域的软骨细胞代谢至关重要。为此，我们采用激光捕获显微切割（LCM）技术来分析病理性软骨和对照软骨不同区域的软骨细胞代谢。在本报告中，我们描述了针对成人人类软骨组织进行LCM的方案。通过该方案，成功地通过LCM获得了软骨组织的特定部位用于基因表达分析。