Hi! I wanted to generate Embryoid bodies(EB's) from mouse embryonic stem cells in a non-Tissue culture treated flask. Now I need to transfer them to a tissue culture treated flask for directed differentiation to neural lineage. I would like to know the protocol for passaging them. Can I use the centrifuge to get the pellet, followed by resuspension and then seed them? or should I do gravity centrifugation? The reason for my question is, If I use the centrifuge and resuspend the pellet, they would no longer be in EB form and I dont think they can be used for differentiation? Since this is the first time I'am doing this, I would like to have some suggestions to proceed with this issue!
Thank you!
Hello
Look at links here , hope help you
Good Luck
https://www.researchgate.net/topic/Embryoid-Bodies
http://www.sciencedirect.com/science/article/pii/S1873506113001074
https://www.stemcell.com/media/files/manual/MA29141-Maintenance_mESCs_miPSCs_Using_ES_Cult.pdf
Hi Anirudh,
these are good references from Alkarim.
In general, EBs will not easily disintegrate once formed and have started differentiating. Usually it is the opposite problem: trouble with gentle single cell dissociation for things like flow cytometry. You can spin them gently (
Hi, I think the fast way is just to pick up them with the pippet wearing yellow tips and transfer therm to gelatin coated dishes
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