Semiquantitative analysis of gene expression in cultured chondrocytes by RT-PCR.
Reverse transcriptase-polymerase chain reaction (RT-PCR) is a powerful, sensitive, and rapid method to monitor small amounts of nucleic acids. This is of particular interest for small amounts of cells, as in cartilage. We present here two protocols to isolate total RNA and a protocol to study matrix metalloproteinase and type II collagen gene expression from chondrocytes of human origin. Specific gene expression is revealed on an ethidium bromide-containing agarose gel on an ultraviolet plate and normalized to that of a housekeeping gene.
Rolland-Valognes
Gaëlle
G
Division der Rhumatologie, Institut de Recherches Servier, Suresnes, France.
D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D024043:Collagen Type II / Q000235:genetics; D017364:Collagenases / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3 / Q000235:genetics; D012313:RNA / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D024043:Collagen Type II; D012313:RNA; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3