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EB formation


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#1 fakeopal

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Posted 17 February 2009 - 06:25 PM

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!

Edited by fakeopal, 17 February 2009 - 06:31 PM.


#2 pcrman

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Posted 18 February 2009 - 04:56 PM

Although you can use petri dishes (for growing bacteria) for EB formation, I doubt you have that in small well format such as 6-well.

For EB formation, are you using the hanging drop method?

#3 fakeopal

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Posted 20 February 2009 - 02:28 PM

Although you can use petri dishes (for growing bacteria) for EB formation, I doubt you have that in small well format such as 6-well.

For EB formation, are you using the hanging drop method?


Hey PCRman,

Thanks for the reply. No I'm growing my EB using suspension method. I finally convinced my lab manager and we are going to have the non-sticky culture dish available. So I'm going to try growing my EB in those. I don't think it's recommended to use those fisherbrand petri dishes. But if anyone is willing to try, let me know how the result turns out =)

Hel




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