Thread: EB formation - which type of culture dish should I use?

I'm working with mouse embryonic stem cells and I have encountered some trouble during EB formation. I just realized that I have been using the wrong kind of tissue culture dish for growing EB -- the one I was using are Corning treated culture dish (product #430167) that are optimal for cell attachment. After differentiation, the cells grow on the bottom of the plate as a monolayer of cells instead of floating cell aggregates. I should really be using Non-treated culture dish or the Ultra-low attachment culture dish. I wanted to purchase those dishes but my lab manager said I can use the petri dish generally used for pouring agar plates (Fisherbrand, product #08-757-13)

Can anyone tell me whether this type of petri dish can be used for culturing ES/EB? Has anyone ever used it for this purpose?

Also, I was wondering if I have over-trypsinized the ES cells off the feeders (I think I let that step go for ~5 minutes last time) that might have caused the failure of cell aggregation. Can someone please help me with this?

Thanks a bunch!

Well, I'm not a specialist in this field but as far as I know, cell cultures are usually grown in so-called mattresses. But I believe they are made from the same plastic so you could use Petri dishes.
If you are worried about the intensity of the tripsine treatment you could try to repeat the experiment using less tripsine or less time for tripsine treatment.