Life Technologies’ complete primary cell solutions enable scientists to investigate these angiogenic processes in a simple and optimized model. These robust tools include cells culture and detection reagents, and validated protocols to facilitate your research. Whether you are new to angiogenesis research and would benefit from a complete kit, or are an experienced scientist and want to pick and choose the reagents that are ideal for your protocol, we have solutions for all your angiogenesis research needs. Learn more about the angiogenesis process >

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Step-by-Step Guide to Angiogenesis Products

Human Umbilical Vein Epithelial Cells

Gibco® primary human umbilical vein endothelial cells (HUVECs) are cryopreserved at the end of the primary culture. Primary HUVECs are prepared to provide ≥70% viability upon thawing, with each vial containing sufficient cells to seed  ~100 cm2 of tissue culture surface (>/=500,000 cells/vial).
Each lot of HUVECs undergoes performance testing and is guaranteed to achieve at least 16 population doublings (PD) after thawing when using Medium 200 with LVES growth supplement. HUVECs must take up aceylated-LDL and express von Willebrand factor (vWf ) and CD31 (endothelial cell markers), but not α-actin (a smooth muscle cell marker).

 

Recommended Cells:

Large Vessel Media Systems

Optimized for use with Gibco® media

Gibco® primary human umbilical vein endothelial cells (HUVECs)  have been optimized for use with our Medium 200 or Medium 200 PRF, and Large Vessel Endothelial Supplement. LVES delivers:

  • Superior growth rates of endothelial cells
  • Maximum number of passages
  • Retained cell characteristics (morphology, biomarker expression, cellular function, etc.)
  • Optimal performance in tube formation, cell invasion, cell migration and cell proliferation assays 

   

Figure 1.  LVES is a complete, ready-to-use blend of growth supplements optimized to deliver >4 x 104 HUVEC per cm2 when used with Medium 200 or Medium 200PRF.  Preparation of supplemented media is a simple process, and the final solution can be stored for up to 30 days at 4°C protected from light.

 

Recommended Media and Supplements:

Basement membrane systems

Geltrex® LDEV-Free Matrix

Geltrex® LDEV-Free Reduced Growth Factor Basement Membrane Matrix is a soluble, reduced growth factor, lactate dehydrogenase-elevating virus-free extract used in a wide variety of cell culture applications for promotion and maintenance of many cell types, including primary epithelial cells, endothelial cells, and stem cells. When used undiluted as a thick gel preparation, it creates a more physiologically relevant environment for angiogenesis assays. It can also be diluted for use as a coating matrix for culture vessels. Consistent protein concentration lot-to-lot helps reduce the need to screen lots.

We have developed a tube formation assay protocol which describes the optimal use of the products in our angiogenesis workflow.  This assay measures the ability of endothelial cells, plated at subconfluent densities on our Geltrex® LDEV-Free Matrix, to form capillary-like structures (a.k.a. tubes).   This assay determines the ability of various components to promote or inhibit tube formation.

Phase contrast

Calcein AM

Figure 2. Endothelial tube formation assay performed using Angiogenesis Starter Kit protocol and reagents: Human Umbilical Vein Endothelial Cells (HUVEC) cultured in Medium 200 (M200) basal medium supplemented with Large Vessel Endothelial Supplement (LVES) seeded onto a Geltrex® thin gel coating .  Representative images following 18 hr incubation, (left) phase contrast image and (right) fluorescent  staining using Calcein AM.  Same field shown in both panels.

 

Recommended Matrices:

Human Umbilical Vein Epithelial Cells

Life Technologies offers a host of products that enable further engineering of primary cells, as well as analysis of activity and function within primary cell models. Molecular Probes® dyes work well for angiogenesis applications, including Calcein AM, Calcein Blue AM and CellTrace™ Calcein Red-Orange, and detection technologies such as CellLight® reagents.

CellLight® reagents are ready-to-use fluorescent protein constructs targeted to specific subcellular structures.  These reagents provide a simple and effective method for introducing targeted intracellular labels within living cells.  Simply add the reagent to your cells, incubate overnight, and you’re ready to image your cells.  CellLight® reagents come in a variety of colors and targets, including actin, endsomes, lysosomes, and tubulin, for convenient multiplexing and co-localization studies.

Cellular labeling with CellLight® reagents employs BacMam technology, which uses a modified insect cell baculovirus coupled with a mammalian promoter as a vehicle to efficiently deliver and express genes in mammalian cells. Unlike expression vectors, BacMam reagents enable titratable and reproducible expression and offer high cotransduction efficiency, enabling multiple BacMam reagents to be used in the same cell.

We have developed a tube formation assay protocol which describes the optimal use of the products in our angiogenesis workflow.  This assay measures the ability of endothelial cells, plated at subconfluent densities on our Geltrex® LDEV-Free Matrix, to form capillary-like structures (a.k.a. tubes).   This assay determines the ability of various components to promote or inhibit tube formation.

 

Recommended Cells:

Resources

Life Technologies provides a comprehensive set of resources for primary cell research, including protocols and technical support.

Use the links (right) to browse our current information on angiogenesis research.

Recommended Recources

Angiogenesis Starter Protocol

HUVEC Tube Formation Image (18 Hours)

About Angiogenesis

Angiogenesis—the formation of new blood vessels from existing vasculature—is an integral part of both normal and pathological processes. It is required for tumor growth and metastatic spread, and as a result is a hot research area within oncology. This complicated process results from the input of multiple signaling pathways.

During angiogenesis, endothelial cells to disrupt the surrounding basement membrane, migrate toward an angiogenic stimulus, proliferate to form the new vessel, and reorganize to create the necessary three dimensional vessel structure. In vitro assays are widely used to study these functions in the presence of either angiogenic or antiangiogenic agents.

For research use only.  Not for use in diagnostic procedures.