I am trying to do a co-culture of fibroblasts and intestinal organoids, with the fibroblasts as a 2D layer on the bottom of a 48 well and the organoids in a matrigel dome above. To set up this co-culture I first grow a 2D layer of fibroblasts on the bottom of the plate and then remove the media and add a matrigel dome with intestinal organoids on top. The problem I am encountering is that the surface of the 48-well with the fibroblasts seems to remain quite wet/slippery, so that when I pipet on the matrigel it completely disperses over the the bottom and to the edges instead of forming a nice dome. Does anybody have an idea how to overcome this issue and get a nice dome?
Perhaps you could add a thin layer of matrigel on top of the fibroblasts before adding the organoid-containing matrigel on top once the first layer has solidified.
Hello, I tried to set the same system up. It is challenging to keep a matrigel dome if the plastic or glass surface is already hydrated. The way I circumvented this problem was to first set the matrigel dome with my organoid lines and the next day deploy the fibroblast in suspension and allow them to attach.
I was able to get perfect domes with a fibroblast layer. However, there were no fibroblasts under the dome, but these migrated days later.
Thanks for the answers Salvador Flores-Torres John Hardy Lockhart ! both sound like good solutions, though John's answer probably works slightly better with my project. Will give them a try!
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