Dear all,
Recently i have a really crazy and brilliant idea to determine if a compound would be able to increase / decrease the motility of a cell type towards another cell type. How should i go about to prove this hypothesis?
Thank you for your kind responses.
Hi Justin, the easiest way I can think of is to use transwell migration chambers.
Plate cell A onto the lower chamber, plate cell B onto the upper chamber. Have both chambers in media + serum. Add your compound to the lower chamber and see if it stimulates migration of cell B across the transwell membrane (using appropriate negative controls).
Dear all, thank you for your kind responses. Dr Stefansson, i have considered using transwell but my concern is that the pores on the transwell may be too small for the cells to migrate over. Will this be of issue? The pore size of my transwell is 0.4 microns and the diameter of the cell upon trypsinization is around 10 microns .
To add to one other the above answers. If you use the transwell assay instead of growing the cells in the bottom well you could try and use conditioned media from the targeted cells.Might be easier. Speaking of pore size. I used endothelial cells, which are quite large, in transwell assay of 0.4µM and they 'migrated' fine. A cell is a bit like a mouse - it can squeeze through really tiny holes :)
Depending on your throughput you can also look at the Dunn Chemotaxic chamber, or this higher throughput version from IncuCyte, which negates teh rollover effect you get with loss of gradient in a transwell.
Follow the Chemotaxis link https://www.essenbioscience.com/en/applications/live-cell-assays/cell-migration-and-invasion/
You'll need access to an IncuCyte though
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