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

被引:40
作者
Gehrsitz, A [1 ]
McKenna, LA [1 ]
Söder, S [1 ]
Kirchner, T [1 ]
Aigner, T [1 ]
机构
[1] Univ Erlangen Nurnberg, Inst Pathol, Cartilage Res Grp, D-91054 Erlangen, Germany
关键词
D O I
10.1016/S0736-0266(00)90028-7
中图分类号
R826.8 [整形外科学]; R782.2 [口腔颌面部整形外科学]; R726.2 [小儿整形外科学]; R62 [整形外科学(修复外科学)];
学科分类号
摘要
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 lessons 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. (C) 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:478 / 481
页数:4
相关论文
共 9 条
[1]   EXTRACTION AND ISOLATION OF MESSENGER-RNA FROM ADULT ARTICULAR-CARTILAGE [J].
ADAMS, ME ;
HUANG, DQ ;
YAO, LY ;
SANDELL, LJ .
ANALYTICAL BIOCHEMISTRY, 1992, 202 (01) :89-95
[2]   ACTIVATION OF COLLAGEN TYPE-II EXPRESSION IN OSTEOARTHRITIC AND RHEUMATOID CARTILAGE [J].
AIGNER, T ;
STOSS, H ;
WESELOH, G ;
ZEILER, G ;
VONDERMARK, K .
VIRCHOWS ARCHIV B-CELL PATHOLOGY INCLUDING MOLECULAR PATHOLOGY, 1992, 62 (06) :337-345
[3]   Suppression of cartilage matrix gene expression in upper zone chondrocytes of osteoarthritic cartilage [J].
Aigner, T ;
Vornehm, SI ;
Zeiler, G ;
Dudhia, J ;
vonderMark, K ;
Bayliss, MT .
ARTHRITIS AND RHEUMATISM, 1997, 40 (03) :562-569
[4]   THE STRUCTURE AND FUNCTION OF CARTILAGE PROTEOGLYCANS [J].
CARNEY, SL ;
MUIR, H .
PHYSIOLOGICAL REVIEWS, 1988, 68 (03) :858-910
[5]   Changes in messenger RNA and protein levels of proteoglycans and link protein in human osteoarthritic cartilage samples [J].
CsSzabo, G ;
Melching, LI ;
Roughley, PJ ;
Glant, TT .
ARTHRITIS AND RHEUMATISM, 1997, 40 (06) :1037-1045
[6]   BIOCHEMICAL AND METABOLIC ABNORMALITIES IN ARTICULAR CARTILAGE FROM OSTEO-ARTHRITIC HUMAN HIPS .2. CORRELATION OF MORPHOLOGY WITH BIOCHEMICAL AND METABOLIC DATA [J].
MANKIN, HJ ;
DORFMAN, H ;
LIPPIELL.L ;
ZARINS, A .
JOURNAL OF BONE AND JOINT SURGERY-AMERICAN VOLUME, 1971, A 53 (03) :523-+
[7]   DISCOORDINATE GENE-EXPRESSION OF AGGRECAN AND TYPE-II COLLAGEN IN EXPERIMENTAL OSTEOARTHRITIS [J].
MATYAS, JR ;
ADAMS, ME ;
HUANG, DQ ;
SANDELL, LJ .
ARTHRITIS AND RHEUMATISM, 1995, 38 (03) :420-425
[8]   Analysis of proteoglycan messages in human articular cartilage by a competitive PCR technique [J].
Melching, LI ;
CsSzabo, G ;
Roughley, PJ .
MATRIX BIOLOGY, 1997, 16 (01) :1-11
[9]   RNA ISOLATION FROM CARTILAGE USING DENSITY GRADIENT CENTRIFUGATION IN CESIUM TRIFLUOROACETATE - AN RNA PREPARATION TECHNIQUE EFFECTIVE IN THE PRESENCE OF HIGH PROTEOGLYCAN CONTENT [J].
SMALE, G ;
SASSE, J .
ANALYTICAL BIOCHEMISTRY, 1992, 203 (02) :352-356