Bilge Alptekin, İbrahim Alptekin, Ezel Erkan, Ferda Topal Celikkan, Alp Can
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引用次数: 0
Abstract
This chapter presents a stepwise optimization of a Matrigel®-based in vitro angiogenesis assay using human umbilical vein endothelial cells (HUVECs) to achieve reproducible and quantifiable tube formation. Critical parameters-including culture surface, matrix composition and thickness, coating temperature and incubation time, growth factor-defined serum-free medium, and cell seeding density-were systematically evaluated. Optimal conditions were identified as 3 mg/mL Matrigel (100 μL/well), polymerized at 22 °C for 90 min, with HUVECs seeded at 20,000 cells/cm2 and cultured in a defined growth factor cocktail. Standardized imaging and ImageJ-based analysis enabled robust quantification of network features. This protocol improves consistency, stability, and comparability across angiogenesis studies.
期刊介绍：
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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