My cells were transfected with Wnt5A containing vector and stably express Wnt5A. In scratch assay, these cells show increased migration relative to mock but in transwell migration assay ,their migration and invasion is lower than mock cells.Is there a specific reason for this?
In scratch assay usually you see only migration, and in transwell migration assay there is also an effect of cell invasion through the filter hollows. Personally, I would suggest 3D inverse invasion assay, when the effect of gravity is eliminated, and cell are migrating up to the attractant. Then you may distinguish both effects and measure them separately. If you want to see only the effect on migration, it should be better to use microfluidic chamber with the gradient of attractant, as in scratch assay the cells are pushing each other and this assay is not very "pure"migration assay.
Migration and invasion both are different properties of a cell. May be your cells are showing only migration as observed by scratch assay. MMP secretion is required to go through from the matrix for the invasion.
I agree with Vilma. Your cells with enhanced Wnt5A might only have effects on migration but not invasion. Compared to in vitro cell culture, you can use mice to see the in vivo effects of increased Wnt5A. After all, the differences between them are huge.
Besides, you can also detect some pathways related to migration and invasion, i.e., FAK pathway. Some chemokines can also be measured as well.
Good luck!
Jennifer
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