Analysis of in vivo endothelial cell activation applying RT-PCR following endothelial cell isolation by laser dissection microscopy.
Laser dissection microscopy was applied to isolate endothelial cells from tumors obtained from mice treated with TNF-alpha. RNA integrity was demonstrated from whole sections and dissected cells after acetone fixation and hematoxylin staining. RT-PCR for GAPDH, CD31, VCAM-1, ICAM-1, and E-selectin was successfully performed on these samples. These data demonstrate the feasibility of analyzing local endothelial cell responses in diseased tissues at the level of gene expression.
Asgeirsdottir
S A
SA
Department of Pharmacokinetics and Drug Delivery, Medical Biology Section, Groningen University Institute for Drug Exploration (GUIDE), Groningen, the Netherlands. s.asgeirsdottir@farm.rug.nl
Werner
N
N
Harms
G
G
Van Den Berg
A
A
Molema
G
G
D000818:Animals; D019040:E-Selectin / Q000235:genetics; D004730:Endothelium, Vascular / Q000473:pathology / Q000503:physiopathology*; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D007834:Lasers; D051379:Mice; D008853:Microscopy; D009374:Neoplasms, Experimental / Q000098:blood supply / Q000235:genetics; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics
D019040:E-Selectin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1