Hi everyone,
I started using Matrigel about 6 months ago for my cell invasion assays. The first batch (for which I didn't write down the lot number) gave me great results (upper row in the picture). The spheroids were formed nicely and the cells could survive and invade after 7 days of starting the assay.
Then we bought another bottle of Matrigel and from then on I couldn't get the same nicely-formed spheroids (lower row). Even the invading cells look different. Before they were these snake-like protrusions from the spheroid, but now it just looks like the spheroids are falling apart and the outer layer of the spheroid looks dead.
Since I don't know the lot number of the first batch, I don't know the concentration of the protein that I've used. Has anybody else encountered this problem? should I increase/decrease matrigel concentration?
Matrigel growth factor content can vary from lot to lot. It is more likely a Matrigel growth factor problem than a extracellular matrix concentration problem. You need to test each batch. Could you try a purified collagen/laminin mix instead?
Hi Ellen A G Chernoff . Thank you so much for your answer!
I wasn't aware that GF content varies from batch to batch. Any recommendations for dealing with this particular problem (e.g. would increasing the concentration of matrigel actually solve this problem)?
And yes, the next step I planned was to test it in purified collagen. If you have any recommendations/protocols for this as well, I would highly appreciate it.
You will just have to try the increased concentration to see if it helps.
There is a "reduced growth factor" matrigel. See Corning website. https://www.corning.com/catalog/cls/documents/faqs/CLS-DL-CC-026.pdf
to embed in plain Type I collagem, here is a generalized protocol with a suggested collagen concentration: https://engineering.virginia.edu/sites/default/files/common/faculty_groups/lazzara_group/protocols/Growing%20cell%20spheroids%20in%20collagen.pdf
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