AI Article Synopsis
- Angiogenesis is important for both normal body functions and diseases, notably tumor growth and eye disorders, making it vital to understand its molecular mechanisms.
- The study introduces in vitro assays, specifically using human umbilical vein endothelial cells (HUVECs), to model angiogenesis and identify therapeutic targets.
- It details two types of assays: a scratch wound migration assay for 2D cell migration and a spheroid sprouting assay for 3D cell behavior, along with strategies for sample preparation for further molecular studies.
Article Abstract
Angiogenesis plays a crucial role in both physiological and pathological processes within the body including tumor growth or neovascular eye disease. A detailed understanding of the underlying molecular mechanisms and reliable screening models are essential for targeting diseases effectively and developing new therapeutic options. Several in vitro assays have been developed to model angiogenesis, capitalizing on the opportunities a controlled environment provides to elucidate angiogenic drivers at a molecular level and screen for therapeutic targets. This study presents workflows for investigating angiogenesis in vitro using human umbilical vein endothelial cells (HUVECs). We detail a scratch wound migration assay utilizing a live cell imaging system measuring endothelial cell migration in a 2D setting and the spheroid sprouting assay assessing endothelial cell sprouting in a 3D setting provided by a collagen matrix. Additionally, we outline strategies for sample preparation to enable further molecular analyses such as transcriptomics, particularly in the 3D setting, including RNA extraction as well as immunocytochemistry. Altogether, this framework offers scientists a reliable and versatile toolset to pursue their scientific inquiries in in vitro angiogenesis assays.
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