soft agar assay - (Feb/27/2008 )
we like to screen transformed candidates for subcutaneous xenografting; we think of a soft agar assay to check motility; but how to perform?
is it possible to perform with collagen to check invasiveness?
Although I have never done these assays, I don't think soft agar is a good motility assay as it's difficult to follow individual cells. Generally I see it used to show cells don't require substrate adhesion, a hallmark of transformed cells. Check out this paper at pubmed (PMID: 16266993 [PubMed - indexed for MEDLINE]) or let me know if you can't get to it and I'll send it. This group does a very simple assay for motility whereby they grow cells to confluency and then scratch a line away with a sterile pipette and monitor for the ability of cells to migrate into this cleared off area. It sounds pretty easy and straight-forward. Usually a good place to start experiments. Again, I've never actually done these assays but I instantly thought of this paper and thought you might find it to be of some help.
wouldn't it be easier to use a boyden chamber?
thanks vetticus, we use Corning´s Transwell plates and Video microscopy but we thought of an additional kind of motility assay..
yes, rkay, thank you, scratch wound assays show also motility, however, the simple version of hand-made scratches is difficult to perform...