VitroGel® Angiogenesis Assay Kit

ready-to-use, hydrogel system for 2D gel coating and 3D culture of angiogenesis tube formation, invasion, and animal injection.

VitroGel Angiogenesis Assay Kit

  • Xeno-free hydrogel system supporting the angiogenesis process
  • Multiple applications in one kit:  Tube formation, invasion, and animal injection
  • Control the hydrogel properties:  Add your own growth factors and compare with positive control
  • Easy, efficient, and enzyme-free cell harvesting in 20 minutes

VitroGel Angiogenesis Assay Kit is a revolutionary tool for researchers to study the effect of both hydrogel properties and culture medium on the angiogenesis process. The kit can be used to study the angiogenesis tube formation and invasion on both 2D hydrogel coating method and 3D cell culture method. The VitroGel system is also good for animal injection for in vivo study.

Angiogenesis is a highly regulated process that involves the growth of new blood vessels from the existing vasculature. This process plays an important role in both normal developmental processes and numerous pathologies, including wound healing, tumor growth, and metastasis to inflammation and ocular disease.

Traditional angiogenesis assay highly relies on natural extracellular matrix (natural ECM), which has non-adjustable hydrogel compositions and properties.  Therefore, our understanding of the angiogenesis process is limited by studying the molecular cues such as growth factors and inhibitors in culture medium only. There is a lack of knowledge on how hydrogel properties affect the angiogenesis process.

There are two versions of VitroGel Angiogenesis Assay Kits:

  • VitroGel Angiogenesis Assay Kit (Ready-To-Use):  Ready-to-use with a fixed hydrogel mechanical strength to support the angiogenesis assay with adjustable supplements
  • VitroGel Angiogenesis Assay HC Kit (Cat No. TWG011-K):  Assay kit with a tunable high-concentration hydrogel to allow full control of the hydrogel’s mechanical strength with adjustable supplements.

Depending on the kit type, the ready-to-use VitroGel Angiogenesis Assay Kits can contain the following components:

  • VitroGel Hydrogel Matrix, a xeno-free ready-to-use hydrogel.
  • AAK Supplement 1, a hydrogel supplement without vascular endothelial growth factors (VEGFs) for cell attachment and growth.
  • AAK Supplement 2, a hydrogel supplement with VEGFs to promote tube formation and as a positive control.

All the components can be purchased separately.
Additional AAK Supplements can be purchased separately here >

The VitroGel Hydrogel Matrix is room temperature stable and can be directly mixed with each supplement at 2:1 (v/v) ratio for hydrogel formation.  Researchers can adjust the hydrogel’s molecular cues by adding the growth factors/inhibitors directly to the supplements before mixing with the VitroGel Hydrogel Matrix hydrogel.  Cells cultured in this system can be harvested further easily with the VitroGel Cell Recovery Solution.

2D Hydrogel Coating for Tube Formation and Invasion Workflow

3D Cell Culture Workflow

Specifications

TYPE 1 Kit Contents
  • VitroGel Hydrogel (2 mL)
  • AAK Supplement 1 (1 x 500 µL)
  • AAK Supplement 2 (1 x 500 µL)
  • TYPE 2 Kit Contents
  • VitroGel Hydrogel (2 mL)
  • AAK Supplement 1 (2 x 500 µL)
  • TYPE 3 Kit Contents
  • VitroGel Hydrogel (2 mL)
  • AAK Supplement 2 (2 x 500 µL)
  • Formulation
  • Ready-to-use, xeno-free functional hydrogel
  • AAK Supplement 1: Without VEGFs
  • AAK Supplement 2: With VEGFs
  • UseAngiogenesis Assay, tube formation, invasion, animal injection
    BiocompatibilityBiocompatible, safe for animal studies
    InjectionInjectable hydrogel for in vivo studies and lab automation
    Cell HarvestingVitroGel Organoid Recovery Solution
    5-15 min cell recovery
    pHNeutral
    ShippingSupplements require dry ice shipment.
    Storage
  • VitroGel Hydrogel: store at 2-8°C
  • AAK Supplement 1 and AAK Supplement 2: store at -20°C
  • Number of Uses60 uses at 50 µL per well

    Recommended Product

    Recover cells from the hydrogel within 15 minutes with high cell viability. Non-enzymatic system.

    VitroGel Organoid Recovery Solution

    Protocols / Handbooks / Resources

    Video Protocols & Demonstrations

    VIDEO PROTOCOL TIP

    3D Cell Culture
    “Ready-To-Use” Hydrogels

    VIDEO PROTOCOL TIP

    2D Coating Culture
    “Ready-To-Use” Hydrogels

    VIDEO PROTOCOL TIP

    Cell Recovery
    from 3D & 2D

    VIDEO PROTOCOL TIP

    Injectable Hydrogel
    VitroGel System

    Application Notes

    APPLICATION NOTE

    Xeno-Free Angiogenesis Cell Models Using the VitroGel System

    Data and References

    Figure 1.  Tube formation of endothelial cells on top of VitroGel AAK hydrogel with tube formation supplement, AAK Supplement 2.
    The time-lapse video above shows the tube formation during 18 hours after Human Umbilical Vein Endothelial Cells (HUVEC) seeding on top of VitroGel AAK hydrogel with AAK Supplement 2. The image above shows the tube morphology of HUVEC cells on top of VitroGel AAK hydrogel.  The cells were fixed and stained with DAPI (blue) and ActinGreen™ (green).

    Figure 2. HUVEC cell growth on top of VitroGel AAK hydrogel with cell growth supplement, AAK Supplement 1.
    The image above shows HUVEC cells attached and growing on the surface of VitroGel AAK hydrogel with cell growth supplement, AAK Supplement 1. The cells were fixed and stained with DRAQ5™ (red) and ActinGeen™ (green).

    Figure 3. Comparison of the growth of endothelial cells on top of VitroGel AAK hydrogel with and without growth factor supplement.
    Top Row)
    The time-lapse video shows the tube formation during 18 hours after Human Umbilical Vein Endothelial Cells (HUVEC) seeding on top of VitroGel AAK hydrogel with AAK Supplement 2 containing VEGFs. The image shows the cells attached on the surface of the hydrogel forming luminal structures after 8 hours, which further developed into a tube structures (red arrows). Bottom Row) HUVEC cells grow on the top of VitroGel AAK hydrogel without the cell growth factors. The time-lapse video shows the poor cell attachment and tube formation when there is a lack of growth factors in the hydrogel matrix. The images show the cells attached on the surface of the hydrogel forming less tube structures (red arrows) than cells with full growth factor supplement.

     

    Figure 4. 3D tube structure of HUVEC cells cultured within the VitroGel Angiogenesis Assay Kit. HUVEC cells were mixed with VitroGel AAK hydrogel supplement with tube formation growth factors (AAK Supplement 2) and cultured for 7 days. The video shows the z-stack images of the 3D tube structure formed within the hydrogel. The image above shows the tube networking structure within the VitroGel AAK hydrogel.

    References/Publications

    AAK Kit

    TYPE 1, TYPE 2, TYPE 3, Hydrogel