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Cell Biologics' products can be purchased through Fisher Scientic by searching Cell Biologics' product catalog number or key words...

Meet Us

Harvard Medical School Feb. 1, 2018 - MA

Experimental Biology April 22-24, 2018 - San Diego

PROMOTIONS


  • Get Free Gelatin-Based Coating Solution per primary cell order,  which can be used for most primary cell culture from Cell Biologics through 2018.

NEW PRODUCTS


 

Endothelial Tube Formation Assay

 

Mouse Endothelial Tube Formation Assay (In Vitro Angiogenesis)


Preparing Matrigel:

  • Thaw reduced growth factor matrigel (BD: 354234) by submerging the bottle on ice and store the matrigel at 4 °C overnight.

Preparing Cells:

·       Wash 90-95% confluent endothelial cells with sterile PBS (1X) without calcium and magnesium twice.

·       Incubate cells (on T25 or T75flask) with warm 0.25% Trypsin-EDTA solution (Cell Biologics, Catalog No. 6914) for 3-5 minutes at 37°C. Once cells detach (slightly taps on the flask can be used to speed up the detachment), add 6-10 ml culture medium (Cell Biologics' Cell Culture Medium supplemented with 5-10 % FBS) to the flask to end the digestion (neutralizing the Trypsin).

·       Filter the cell suspension through a sterile 40µm nylon mesh filter (BD 352340) to remove remaining clumps.

·       Count cells and resuspend cells to a final concentration of 2.5-5.0 x 105 cells/ml in endothelial cell culture medium with 50 ng/ml VEGF.


Tube Formation Assay

·       Pipet 200 µl matrigel into each well of a 24-well plate. Avoid creating bubbles.

·       Incubate the 24-well plate at 37 °C and 5% CO2 for 30 min to solidify matrigel.

·       Once matrigel has set, seed 50,000 cells -100,000(200ul) into each well.

·       Observe the tube formation after 4-6 hrs. Peak tube formation may occur between 12hrs and 24hrs. 

·       Take image by 4X lenses microscope. 

Note: Adjust the volume of endothelial culture medium and number of cells plated depending on the type of endothelial cell used.