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?

-The Bearer-

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.

-rkay447-

wouldn't it be easier to use a boyden chamber?

V

-vetticus3-

QUOTE (vetticus3 @ Feb 27 2008, 11:00 PM)

wouldn't it be easier to use a boyden chamber?

V

thanks vetticus, we use Corning´s Transwell plates and Video microscopy but we thought of an additional kind of motility assay..

-The Bearer-

QUOTE (rkay447 @ Feb 27 2008, 05:05 PM)

yes, rkay, thank you, scratch wound assays show also motility, however, the simple version of hand-made scratches is difficult to perform...

-The Bearer-