Cultrex® In Vitro Angiogenesis Assay Tube Formation Kit

Angiogenesis assay human - hRMVEC

Experiment
Angiogenesis assay human - hRMVEC
Product
Cultrex® In Vitro Angiogenesis Assay Tube Formation Kit from Bio-Techne
Manufacturer
Bio-Techne

Protocol tips

Protocol tips
- Cells are coated with reduced growth factor basement membrane extract (RGF-BME).

Publication protocol

In vitro angiogenesis assay was performed to evaluate the effect of honokiol on the ability of hRMVECs to form tubes on basement membrane, under the trigger of pro-angiogenic factors (VEGF). To carry out this assay, in vitro angiogenesis tube formation kit was used (Trevigen, Gaithersburg, MD). A 96-well plate was quoted with 50 µL of reduced growth factor basement membrane extract (RGF-BME). RGF-BME was allowed to solidify by incubating at 37°C for 1 h. hRMVECs were plated slowly on the membrane at a density of 3×104 cells/well. The cells were treated with different treatment conditions. Cells in VEGF containing ENDO-growth media served as the positive control. The two negative controls used in this experiment were cells incubated in ENDO-basal media and ENDO-growth media with 100 µM sulforaphane. In order to evaluate the effect of honokiol on the tube formation, different concentrations of honokiol (2.5–40 µM) were added in the ENDO-growth media. Cells were incubated for 18 h at 37°C in a CO2 incubator. The media was carefully aspirated and cells were washed with 150 µL PBS. Further to each well 2 µM Calcein AM (in PBS) was added and incubated for 30 min at 37°C. After the incubation Calcein AM was removed and PBS was added. Extent of tube formation was assayed using fluorescence images. Efficacy of honokiol on tube formation was evaluated by quantitating the average branch length using angiogenesis analyzer plugin for ImageJ.


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Discussion

Discussion

5 years ago

Author: Sander J. Mikkelsen Denmark

endothelial cells vs HUVEC cells?

What is the difference between endothelial cells and HUVEC cells for angiogenesis assay?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Angiogenesis assay human - hRMVEC using Cultrex® In Vitro Angiogenesis Assay Tube Formation Kit from Bio-Techne.

Paper title
Evaluation of Anti-HIF and Anti-Angiogenic Properties of Honokiol for the Treatment of Ocular Neovascular Diseases
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Manufacturer protocol

Download the product protocol from Bio-Techne for Cultrex® In Vitro Angiogenesis Assay Tube Formation Kit below.

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