I have used BD invasion chambers before on WM9 metastatic melanoma cell and they worked well showing lots of invasion for the controls (repeated 3 times). Now using the same cells and methods the controls aren't invading. Would anyone know why?
Are you using the same batch of cells or did you thaw new ones - there might be a certain variation in different batches of the same cell line?
Also it might be good to include more than one positive controll cell line for your invasion-assays.
Different batches of trypsin used to detach cells prior to your assay could also have an impact. If your new trypsin is e.g. more active it could stress your cells more than usual and/or remove additional proteins (receptors..) from the cell membrane thereby reducing the invasion phenotype. Maybe detaching cells non-enzymatically (simply try 1 to 10mM EDTA for some minutes) would be better in your case?
Best regards, Christian
The cells were different passages from the same batch, but I have opened a new bottle of trypsin recently. I'll try using non-enzymatic detachment.
Thanks for the suggestion
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