How do I reduce spontaneous formation of tubular structures on Cultrex BME in the absence of angiogenic factors?

Primary endothelial cells, such as Human Umbilical Vein Endothelial Cells (HUVECs) form capillary-like structures in the absence of added angiogenic factors less often than immortalized endothelial cells. Generally, reducing the number of cells per cm2 plated onto Cultrex BME will result in less background or spontaneous tube formation. Titrate the number of cells and find optimal conditions for your specific cell line. When endothelial cells fully form capillary structures in response to angiogenic activators, but not in their absence, you may proceed.

What are the advantages of the Tube Formation Assay?

The Tube Formation Assay is the most widely used in vitro angiogenesis assay. The assay is rapid, inexpensive and quantifiable. It can be used to identify potentially angiogenic and anti-angiogenic factors, to determine endothelial cell phenotype, and to study pathways and mechanisms involved in angiogenesis. It can be performed in a high throughput mode to screen for a large number of compounds.

What are the variables associated with the Tube Formation Assay?

The major variables associated with tube formation are composition of the Cultrex Basement Membrane Extract (BME) hydrogel, thickness of the hydrogel, cell density, composition of angiogenic factors in the assay medium, and assay period.

What cell types can be used in the Tube Formation Assay?

The Tube Formation Assay is specific for endothelial cells, either primary cells or immortalized cell lines. Only endothelial cells form capillary-like structures with a lumen inside. Other endothelial cell types form other structures.

What is the Tube Formation Assay?

The Tube Formation Assay is based on the ability of endothelial cells to form three-dimensional capillary-like tubular structures when cultured on a hydrogel of reconstituted basement membrane, such as Cultrex Basement Membrane Extract (BME).

Which Cultrex Basement Membrane Extract (BME) should I use for the Tube Formation Assay?

Cultrex Reduced Growth Factor BME (RGF BME) is generally used for testing compounds that promote angiogenesis because formation of capillary-like structures (tubes) is significantly less compared to non-growth factor reduced varieties of Cultrex BME. The Cultrex In Vitro Angiogeneis Assay (Tube Formation) includes a qualified production lot of Cultrex RGF BME that exhibits reduced background tube formation in the absence of angiogenic factors.

Cultrex In Vitro Angiogenesis Assay (Tube Formation)

Name: Cultrex In Vitro Angiogenesis Assay (Tube Formation)
Brand: R&D Systems
SKU: 3470-096-K
Rating: 5 (4 reviews)

R&D Systems, part of Bio-Techne | Catalog # 3470-096-K

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Product Datasheet / COA

Tube formation assay for evaluation of angiogenesis

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Product Summary
Kit Contents
Preparation & Storage
Background
Product Documents

Key Product Details

Features

Allows for the detection of inducers and inhibitors of endothelial cell tube formation.

Species

Multi-Species

Product Summary for Cultrex In Vitro Angiogenesis Assay (Tube Formation)

Why Use Cultrex In Vitro Angiogenesis Assay (Tube Formation), 96-well?

The Cultrex in vitro Angiogenesis Assay Tube Formation Kit was developed for the evaluation of inducers and inhibitors of endothelial cell tube formation. During the course of the assay, endothelial cells are seeded on a thick gel of Basement Membrane Extract in the presence of angiogenesis modulating agents. If the medium is permissive to the angiogenic response, cells will self assemble into tube networks within hours; if there are inhibitors in the medium, tube network formation will be incomplete or nonexistent. Sulforaphane [1- isothiocyanato-(4R)- methylsulfinyl)-butane], found in broccoli and other cruciferous vegetables, is a naturally occurring cancer chemopreventive agent, and is provided as a control inhibitor to visualize this process. Endothelial cells may be fluorescently labeled using Calcein AM, and images captured using a microscope camera may be analyze to quantify the angiogenic response. Cultrex in vitro Angiogenesis Assay Kit provides a cost-effective method for investigation of prospective angiogenesis inhibitors in a 96 well plate format.

Kit Contents for Cultrex In Vitro Angiogenesis Assay (Tube Formation)

Cell Staining Solution
Cultrex BME, reduced growth factor without phenol red
Sulforaphane
Calcein AM

Note: The components for this kit may require different storage/shipping temperatures and may arrive in separate packaging.

Formulation, Preparation, and Storage

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended on the product label.

Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Angiogenesis Assay Kits

Angiogenesis is the tightly regulated process by which new blood vessels are formed from the existing vasculature. This process is physiologically important for development and wound healing, and is also a common driver in multiple diseases including rheumatoid arthritis, atherosclerosis, macular degeneration, and cancer. Angiogenesis occurs in response to a variety of molecular cues. Generally, the angiogenic process includes endothelial cell proliferation, chemotactic endothelial cell migration through the extracellular matrix barrier, and the formation of capillary tubes. Physiological and pathological angiogenesis utilize many of the same cellular processes and molecular signaling networks, however the structures that form during pathological angiogenesis are often functionally abnormal.

Cultrex In Vitro Angiogenesis Assay (Tube Formation)

R&D Systems, part of Bio-Techne | Catalog # 3470-096-K

Key Product Details

Features

Species

Product Summary for Cultrex In Vitro Angiogenesis Assay (Tube Formation)

Kit Contents for Cultrex In Vitro Angiogenesis Assay (Tube Formation)

Formulation, Preparation, and Storage

Shipping

Storage

Background: Angiogenesis Assay Kits

Additional Angiogenesis Assay Kits Products

Product Documents for Cultrex In Vitro Angiogenesis Assay (Tube Formation)

Product Datasheets

COA

Certificate of Analysis

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