An in vitro model for studying vascular injury after laser microdissection.
We have developed an in vitro model for studying vascular injury. After 7-10 days in a three-dimensional collagen gel culture, capillary-like tubes were formed in the collagen gels. We injured these capillary-like tubes with a laser microdissection system or a scrape method with razors and then examined the collagen gel culture by phase contrast and electron microscopy. After laser injury, profuse necrotic cells were observed around the injured capillary-like tubes and within the tubular lumen compared to the razor injury. We then isolated total RNA from these cultures and prepared cDNA for investigations by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Quantitative real time RT-PCR revealed the up-regulation of transcription factor early growth response-1 (Egr-1) after both laser and razor injury, accompanied by the up-regulation of fibroblast growth factor-2 (FGF-2), a proangiogenic factor downstream of Egr-1. The effective laser energy is concentrated on the minute focal spot only. These methods provide a useful in vitro model for studying vascular injury.
Fujita
Keiko
K
Department of Anatomy, Saitama Medical School, 38 Moroyama, Iruma-gun, 350-0495, Saitama, Japan. kfujita@saitama-med.ac.jp
Komatsu
Kumiko
K
Tanaka
Kayoko
K
Ohshima
Susumu
S
Asami
Yoshiko
Y
Murata
Eiko
E
Akita
Masumi
M
D000818:Animals; D051766:Early Growth Response Protein 1 / Q000096:biosynthesis; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis; D007834:Lasers; D008297:Male; D051379:Mice; D042282:Microdissection; D008854:Microscopy, Electron; D023421:Models, Animal; D018919:Neovascularization, Physiologic / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D014652:Vascular Diseases / Q000473:pathology*
D051766:Early Growth Response Protein 1; C486109:Egr1 protein, mouse; D016222:Fibroblast Growth Factor 2