An in vitro model for studying vascular injury after laser microdissection
An in vitro model for studying vascular injury after laser microdissection
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DOI：
10.1007/s00418-005-0106-9
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发表时间：
2006-05-01
影响因子：
2.3
通讯作者：
Akita, M
中科院分区：
文献类型：
--
作者：
Fujita, K;Komatsu, K;Akita, M
我们已经开发了一种用于研究血管损伤的体外模型。在三维胶原凝胶培养7-10天后，在胶原凝胶中形成毛细血管样管。我们用激光显微切割系统或剃刀刮伤这些毛细血管样管，然后用相差和电子显微镜检查胶原凝胶培养物。激光损伤后，与剃刀损伤相比，在损伤的毛细血管样管周围和管腔内观察到大量坏死细胞。然后，我们从这些培养物中分离总RNA，并通过定量实时逆转录聚合酶链反应（RT-PCR）制备cDNA用于调查。定量真实的时间RT-PCR揭示了转录因子早期生长反应-1（Egr-1）在激光和剃刀损伤后的上调，伴随着成纤维细胞生长因子-2（FGF-2）的上调，FGF-2是Egr-1下游的促血管生成因子。有效激光能量仅集中在微小焦点上。这些方法为研究血管损伤提供了一个有用的体外模型。
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.