IncuCyte® Cell Migration Kit

Protocol tips

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	- Cells are cultured/seeded in presence of EGM2S + for 18h.. - After cells are carefully washed, medium was then replaced with EGM2S +, EGM2S −, CMS, CMS + Dex (50 nM) or CMD

Protocol tips
- Cells are cultured/seeded in presence of EGM2S + for 18h.. - After cells are carefully washed, medium was then replaced with EGM2S +, EGM2S −, CMS, CMS + Dex (50 nM) or CMD

Publication protocol

HUVEC migration was assessed using the IncuCyte ZOOM Scratch assay (Essen Bioscience, Hertfordshire, UK) according to manufacturer's instructions. Briefly, 3 × 104 HUVECs/well were seeded in 96-well culture plates and cultured for 18 h in EGM2S + at 37 °C, 5% CO2. A scratch was then made using the WoundMaker tool (Essen Bioscience). The cells were washed twice with medium, and the medium was then replaced with EGM2S +, EGM2S −, CMS, CMS + Dex (50 nM) or CMD. 10-Fold concentrated CM were diluted 1:1 with EGM2S −. Plates were then installed in the IncuCyte ZOOM system and images (10 × magnification) of the wound were recorded in each well every hour for 48 h. Scratch closure rate was evaluated with the IncuCyte software, expressed as percentage of relative wound density (RWD) over a 30 h period. RWD = 0 at time 0 and 100% when cell confluence within the wound area is equal to that outside the initial wound area, thus normalizing for changes in cell density due to proliferation or other non-motogenic pharmacological effects. The area under the curve (AUC) was calculated for each condition and the results are expressed as AUC from RWD.


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Manufacturer protocol

Download the product protocol from Essen Bioscience for IncuCyte® Cell Migration Kit below.

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