As an expert in the field of therapeutic antibody development, Creative Biolabs offers various in vitro functional assays to assess antibody bioactivity and to investigate its mechanism of action. Particularly, we offer the endothelial cell tube formation assay described here to screen antibodies with anti-angiogenic potential.
Angiogenesis, the physiological formation of new blood vessels from existing ones, is vital for various processes including organ growth, embryonic development, and wound healing. However, angiogenesis is associated with various pathological conditions, including cancer, psoriasis, diabetic retinopathy, arthritis, asthma, autoimmune disorders, infectious diseases, and atherosclerosis. Physiological angiogenesis is a highly organized process, comprising different steps: basement membrane disruption, endothelial cell migration, proliferation and differentiation into capillaries (Fig.1). One key step of this process is the assembly of endothelial cells into tubes, termed tube formation.
Fig.1 Progression of Angiogenesis. (Fan, 2006)
The tube formation assay is a simple in vitro angiogenesis assay that is routinely used to screen pro-angiogenic or anti-angiogenic potential of substances and compounds. Besides, it can be used to determine genes or pathways involved in angiogenesis. Thus, it is of great utility in diseases like cancer where inhibition of angiogenesis is one of the key research areas to combat the disease. The principles underlying the assay are that endothelial cells retain the ability to divide and migrate rapidly in response to angiogenic signals. When the endothelial cells are cultured on a matrix of basement membrane extract, they can be induced by angiogenic stimuli to differentiate and form tube-like structures within hours. Quantification can be performed by measuring the tube area or the length and/or the number of branch points.
The tube-forming capacity and their response to angiogenesis stimulators vary among different groups of endothelial cells. At Creative Biolabs, we generally use the most commonly used HUVECs. Additionally, we can also use other endothelial cell types in this assay upon requests, such as HDMEC, HCAEC, HPMEX, and HCMEC. We can assist you in the selection of the most appropriate endothelial cell sources.
The extracellular matrix provides a physiological substrate that supports key cellular functions. It plays a role in the structural organization of cells and tissue, cell attachment and proliferation, as well as induction of cell differentiation. Thus, the selection of support matrices is also crucial to obtaining consistent and reliable data.
The in vitro tube formation assay closely mimics the in vivo environment and is thus a powerful tool for checking angiogenesis promoters and inhibitors before using them in in vivo assays. As a leader who has been devoted to therapeutic antibody development and function analysis, Creative Biolabs takes pride in having earned a reputation for accomplishing thousands of challenging projects. Our scientists are willing to custom-design the studies to meet your specific expectations. For more detailed information, please feel free to contact us or directly sent us an inquiry.
Reference
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