使用薄层分析法体外评估内皮细胞的血管生成潜力。

Use of a Thin Layer Assay for Assessing the Angiogenic Potential of Endothelial Cells In Vitro.

作者信息

Lane James A E, Faulkner Ashton, Finding Elizabeth J T, Lynam Eleanor G, Wheeler-Jones Caroline P D

机构信息

Department of Comparative Biomedical Sciences, Royal Veterinary College, London, UK.

Experimental Cardiovascular Medicine, Bristol Medical School, University of Bristol, Bristol, UK.

出版信息

Methods Mol Biol. 2022;2475:197-204. doi: 10.1007/978-1-0716-2217-9_13.

DOI:10.1007/978-1-0716-2217-9_13

PMID:35451758

Abstract

Angiogenesis is essential for wound healing and regeneration and plays a significant role in several pathologies including cancer and atherosclerosis. In vitro assays offer simple and powerful tools for investigating the regulation of the angiogenic functions of primary endothelial cells (ECs) before moving to in vivo studies. The classic in vitro two-dimensional angiogenesis assay utilizes Basement Membrane Extract (BME) to study the differentiation and sprouting of ECs over a 24-h period. The protocol described here details a thin layer BME adaptation of the angiogenesis assay requiring significantly less BME and carried out in 96-well plates, allowing for a larger data yield at a greatly reduced cost, while maintaining the robustness of an assay used extensively over the past three decades.

摘要

血管生成对于伤口愈合和组织再生至关重要，并且在包括癌症和动脉粥样硬化在内的多种病理过程中发挥着重要作用。在进行体内研究之前，体外实验为研究原代内皮细胞（ECs）血管生成功能的调节提供了简单而强大的工具。经典的体外二维血管生成实验利用基底膜提取物（BME）来研究ECs在24小时内的分化和出芽。此处描述的方案详细介绍了一种血管生成实验的薄层BME改良方法，该方法所需的BME显著减少，且在96孔板中进行，能够以大幅降低的成本获得更多数据，同时保持了过去三十多年来广泛使用的实验方法的稳健性。