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小鼠角膜微袋血管生成试验。

The mouse cornea micropocket angiogenesis assay.

作者信息

Rogers Michael S, Birsner Amy E, D'Amato Robert J

机构信息

The Vascular Biology Program, Children's Hospital Boston, and Department of Ophthalmology, Harvard Medical School, 300 Longwood Ave., Boston, Massachusetts 02115, USA.

出版信息

Nat Protoc. 2007;2(10):2545-50. doi: 10.1038/nprot.2007.368.

Abstract

The mouse corneal micropocket angiogenesis assay uses the avascular cornea as a canvas to study angiogenesis in vivo. Through the use of standardized slow-release pellets, a predictable angiogenic response is generated over the course of 5 d and then quantified. Uniform slow-release pellets are prepared by mixing purified angiogenic growth factors such as basic fibroblast growth factor or vascular endothelial growth factor with sucralfate (a stabilizer) and Hydron (poly-HEMA (poly(2-hydroxyethyl methacrylate)) to allow slow release). This mixture is applied to a mesh that controls unit size and then allowed to harden. A micropocket is surgically created in the mouse cornea and a pellet implanted. Five days later, the area of the cornea overgrown by the angiogenic response is measured using a slit lamp. A skilled investigator can implant and grade 40 eyes in about 2.5 h. The results of the assay are used to assess the ability of potential therapeutic molecules or genetic differences to modulate angiogenesis in vivo.

摘要

小鼠角膜微袋血管生成试验利用无血管的角膜作为研究体内血管生成的平台。通过使用标准化的缓释微丸,在5天的过程中产生可预测的血管生成反应,然后进行量化。通过将纯化的血管生成生长因子(如碱性成纤维细胞生长因子或血管内皮生长因子)与硫糖铝(一种稳定剂)和Hydron(聚甲基丙烯酸羟乙酯)混合以实现缓慢释放,从而制备均匀的缓释微丸。将该混合物应用于控制单位大小的网格上,然后使其硬化。在小鼠角膜上通过手术创建一个微袋并植入一个微丸。五天后,使用裂隙灯测量因血管生成反应而过度生长的角膜面积。一名熟练的研究人员大约可以在2.5小时内植入并分级40只眼睛。该试验的结果用于评估潜在治疗分子或基因差异在体内调节血管生成的能力。

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