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Spheroid-Based in vivo Angiogenesis Assay |
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The test model
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In vivo models for rapid and objective testing of anti- or pro-angiogenic compounds are rare. Grafting human endothelial cells that establish a human vasculature in mice is unique[1]. ProQinase has combined these features in its spheroid-based in vivo angiogenesis assay to allow fast in vivo testing and screening of antibodies and compounds.
In order to monitor the efficacy of treatment in live animals human endothelial cells (HUVECs) are transduced by a lentivirus encoding luciferase (HUVECs-LLN). The subcutaneously grafted HUVECs-LLN mature and connect to the mouse vascular system to establish a perfused and pericyte-covered micro vascular bed. The formation of a human vasculature is monitored during therapy by bioluminescence imaging in vivo and ex vivo after plug removal by analysing the luciferase signal.
[1] Alajati, A et al.(2008) Nat Methods. 5(5): 439-45
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Options
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The classical analysis of the graft by histology is also possible. Untransduced HUVECs can be injected in a second plug and analysed at the end of the experiment for microvessel density, coverage and/or perfusion by histology. |
Staining of human ECs (anti-hCD34, red) and mouse
pericytes (anti-SMA, red) in the in vivo Angiogenesis Assay |
Study example
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Compound 1 and antibody 1/2 were tested in the spheroid-based in vivo angiogenesis assay. The human microvessel analysis revealed a strong efficacy for compound 1 and antibody 1 and a moderate effect for antibody 2 compared to the untreated control.
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Our service
Base package
- Generation and implantation of human endothelial cell spheroids (HUVECs-LLN)
- Administration of test compounds (e.g. po, ip, iv) for 21 days
- Testing of Sunitinib as positive control
- Measurement of animal weight
- Bioluminescence imaging (every week)
- Measurement of ex vivo luciferase activity
- Preparation of detailed report
Optional
- Generation and implantation of untransfected human endothelial cell spheroids
- Counting of human blood vessels by human CD34 staining
- Counting of pericyte-coverage by human CD34 and smooth muscle actin staining
- i.v. injection of lectin UEA-l and counting of perfusion by human CD34 and UEA-I staining
Please inquire to receive a custom-tailored quote according to your individual requirements.
Fact sheets
in vivo Angiogenesis Assay - luciferase analysis
in vivo Angiogenesis Assay - classical analysis
Related assays:
Spheroid-Based Cellular Angiogenesis Assay
Cellular Phosphorylation Assays (VEGF-R2, TIE2, EPHB4, PDGFR-beta)
Orthotopic Tumor Models
Subcutaneous Tumor Models / Xenografts
Target-specific Tumor Models with Genetically Engineered Cells
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