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Help needed- EBs were too small for differentiation - (Oct/12/2011 )


My task was to form EB from mouse ipsc (2A-EGFPTg-3F) and from there, someone will use the EBs to differentiate to hematopoeitic cells. But, when we checked it, EBs were formed but they were too small, too small that they couldnt be used for differentiation.

What could have been the cause for this? and how can I improve it?

Thanks a lot!


Feed them regularly.
Which tissue culture plates are you using for EBs formation?

-M Kamran Raja-


I used the round bottom low cell bindin 96 well plate. i seeded 1000 cells in 1row and 3000 cells in the 2nd row. after 2 days, i added 80uL of EB medium. 3 days later i changed the medium.


Try to seed more cells and use larger plates, the cells will clump together as time goes.