Dear colleagues, could you, please, suggest a reliable and affordable protocol, which can be used without changes in parallel to MDA-MB-231 and MCF7 breast cancer cells? We expect that MDA-MB-231 may require something to increase cell-cell interaction, while the MCF7 do not need that.
Basically, we intend to measure the effect of a substance on invasiveness of these cells placed in 3D environment. So, we expect to incubate the spheroids in liquid medium, containing a range of the substance concentrations for 24-72 h and then we intend to graft the spheroids into 3D gel to measure the invasion as maximal invasion distance and invasion area. The gel will be, probably, collagenous, but other options are also interesting for us (the preference now is to use some non-expensive biocompatible and transparent 3d gel, even synthetic as well). Thank you very much.
I think the choice of your 3D gel will be crucial. Depending on the type & contents of your gel, cells will behave differently.
I haven't done many exposures in 3D, but mainly studied MDA-MB-231 motility and invasion in 3D environments. When I inject cells as a spheriod in a 3D collagen gel, MDA231 cells invade as single cells. They behave similarly when suspended as single cells in a 3D Matrigel/collagen mix (not injected as spheriod).
Best of luck,
Thank you, Michiel. Yes, we expect this. I am thinking more about collagen gels as it seems that MDA-MB-231 rely on integrin-mediated motility. I have seen also some papers with spheroid/invasion assays in matrigel/geltrex/cultrex... But the first part is to get spheroids of different cell lines by the same protocol.
This is one of the set-ups we use in the lab:
https://www.ncbi.nlm.nih.gov/pubmed/?term=22018386
Works like a charm, for a lot of cell lines. I would expect MCF7s to be non-invasive and slowly grow / expand the spheroid. MDA231 will already show invading cells 24h after injection, so protocol-wise I would inject cells on day 1 and expose 24h later.
This is another set up we've used in the past, and also works for multiple breast cancer cell lines: https://www.ncbi.nlm.nih.gov/pubmed/?term=25289886
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