How to prevent EB becoming too large
Posted 16 June 2011 - 10:07 AM
I am using mouse ES cells to form floating EB for 8 days to promote neuronal differentiation (Following 2004 Nature Neuroscience Paper, Volume 7 Number 9 Page 1003). My problem is that the floating EB, after 6-8 days in 10 cm petri dish in floating formation, become large and very dense in the center, presumably starting to die.
I do not have shaker in the incubator...Is there a good way to prevent EB becoming too big? Low density? Or shall I split the cells into multiple dishes?
Posted 05 September 2011 - 03:21 AM
Posted 16 October 2011 - 07:36 PM
Posted 16 October 2011 - 07:40 PM
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 and 2 days later, I checked and the result was, theye were too small to be used for differentiation.
I was using mouse ipsc
what did u do?