CytoSelect™ 24-Well Wound Healing Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- After washing at step -7 different stimuli were added (Trx-lps, from S. ratti and T. suis, in concentrations of 3 ng, 30 ng, 300 ng, 1 μg, 10 μg, and 25 μg per 500 μL). As a positive control EGF; 0.5 ng, 5 ng, 10 ng, 15 ng, and 25 ng is added and as a negative control LPS were added.

Protocol tips
- After washing at step -7 different stimuli were added (Trx-lps, from S. ratti and T. suis, in concentrations of 3 ng, 30 ng, 300 ng, 1 μg, 10 μg, and 25 μg per 500 μL). As a positive control EGF; 0.5 ng, 5 ng, 10 ng, 15 ng, and 25 ng is added and as a negative control LPS were added.

Publication protocol

To monitor epithelial cell migration of Caco-2 cells and the ability of Trx-lps to improve the wound healing process, we used the CytoSelect 24-Well Wound Healing Assay (Cell Biolabs, Inc.) according to the manufacturer's instructions. By means of the CytoSelect wound healing inserts a 0.9 mm wound field was generated. 500 μL of a Caco-2 cell suspension (containing 0.5 × 106 cells) was added to each well after the inserts had firm contact with the bottom of the wells. After overnight incubation, a monolayer was formed, the inserts were removed, the cells were washed, and the different stimuli were added. We used both Trx-lps, from S. ratti and T. suis, in concentrations of 3 ng, 30 ng, 300 ng, 1 μg, 10 μg, and 25 μg per 500 μL. As a positive control the human epidermal growth factor (EGF; 0.5 ng, 5 ng, 10 ng, 15 ng, and 25 ng) was included in order to get the proper concentration for maximal wound healing effects. As negative control cell media and LPS were added. An inverted digital microscope (EVOS™ FL Thermo Fisher Scientific) by Advanced Microscopy Group was used for observation (4x magnification). The cells were incubated for 4 days, whereby each 24 h a picture was taken and the percent closure was calculated.


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Manufacturer protocol

Download the product protocol from Cell Biolabs for CytoSelect™ 24-Well Wound Healing Assay below.

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