I'm currently growing Caco-2 cells in T-Flask before transferring them to a Matrigel embedded system. The problem is that obviously these cells don't like the plastic and grow in big groups of cells that eventually cover the whole surface. I was wondering whether plating them on collagen or fibronectin coated surface could make them feel happier and grow faster. Any hints, suggestions or recommendations?
Hi Eric,
I worked with Caco-2 cells for a while and I had no problems with cell clustering. I just used Corning T75 flask without any additional treatment. The secret is to disgreggate the clusters very well, doing "ups & down" with the micropippete before seeding them.
In the only case were I needed to treat the surface was when I used glass coverslips. In that case, poly-L-lysine treatment was enough to make these cells stick to the surface.
Good luck!
Sandra
Eric,
We had a similar problem and found that they dispersed better when plated on collagen I. We also pipet thoroughly before plating. I will add that our Caco-2 cells are very heterogeneous. Some are very large and well spread, whereas others are more colonial. Plating on collagen helped to make the monolayer more uniform in appearance.
Judy
Hello Eric,
For a start you can grow Caco-2 cells in 25cc flask, canted neck. As the cells grow you can sub-culture in 75cc flask inorder to have more cells for your assays.
Kofi
hi Eric,
i have been working with Caco-2 cells so far, i have no problem to culture them on just flasks without any coating. When cells are detached with 0.25 % trypsin -0.05% EDTA for about 10 min, and make them dispersed completely. They would be fine.
Hi Eric, I think you can use the plastic surface treated with ply D-Lysine. Usually all type of cells grow happily on this treated surface. Another matrigel coating is also very good and bring lots of exciting results in other cell type thesedays. In case of fibroncetin and collagen, you have to chek and compare how are they grwoing. May be you cab ceck on different treated suface and compare between them! All the best!!!!
Hi Eric,
our Caco-2 cells also grow in large clusters when seeded on plastic T75, probably similar to yours. However, pipetting them up and down with 500 ml and then 100 ml pipetts helps. If you grow them on Collagen I coated Transwell filters, they tend to form monolayers more rapidly - take TEER measurements to see and compare to uncoated surfaces. I my experience, when cells start to act "funny" test them (possible infection).
Hello Eric,
I have been working with Caco-2 for quite some time now. The parental Caco-2 cell line is very heterogeneous, and starts growing in small aggregates until reaching confluence and forming a monolayer. With continued culture they differentiate and polarize into an enterocytic-like phenotype, forming dome-like structures. You get this type of pattern in any tissue culture treated flask, plate or dish, an even in non-treated coverslips. As previously mentioned, extensive dispersal is detrimental to have a representative cell passage. My experience with matrigel 3D systems is that the process of differentiation and polarization is accelerated. They won`t grow as a monolayer so much, but as individualized and pretty nice cyst-like structures that have been used to study intestinal polarity. I recommend this paper for more info: Gao and Kaestner, 2010, Genes Dev (doi: 10.1101/gad.1921510). Hope it helps!
Best regards,
Bruno
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