I have tried adding initially complete media to coat the surface of the dish but it seems like the cells sometimes do not want to attach. Any suggestions?
Hi Cristina,
How about pre-coating the dishes with gelatin or matrigel?
would it do the trick if I were to pre-coat the dish just with FCS? how long would you normally leave the "coating agent" before platting the cells?
I have never tried the coating with plain FCS or complete medium, but I have the intuition that this might not work as good. The point of precoating is to have some of the extracellular matrix components on the dish to "simulate" the extracellular environment and thus, facilitate adhesion of cells. Usually, gelatin coating is made with 0.1% gelatin/water solution, filter sterilized. Cover the surface of the dishes with enough volume and incuabate 4h at 37º. I guess, o/n incubation can work as well. Then, cool down to 4ºc in the fridge, aspirate the gelatine and add the cell suspension. If you prepare a bunch of dishes, you can preserve them for a week or so in the fridge. If this doesn't work, you could try Matrigel coating, although this is more expensive.
Hi Cristina,
Why do you not try Collagene 1 ? ( 100 µg collagène1 /ml acetic acid 0,1N) let it dry in you fluorodishes overnight. Rinse with complet medium before use.
Of course you can also try Poly-D-Lysine, gelatin
It might depend on your melanoma cell line and definitely you may have problems with primary cultures. So you have to try different methods.
Also in addition to use collagen or matrigel (works fine) you may try to use different brand of fluorodishes (8-well chambers etc). I recommend 8-well chambers slides. They are very efficient, you can try different coating or experimental settings on one glass.
Well as far as i know, there is no trick with growing these melanoma cells. Some of the melanoma cells like SKMEL-2 and SKMEL5 will not for a monolayer at all. So, you can make sure you are using the cell lines that have a property of forming a monolayer.
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