Hello researchers!

I have split my cells from 1 well into 6 wells; the confluency was at 80%. The cells were treated with E8 medium plus ROCK inhibitor (at a 1:1000 ratio from a 10 mM stock solution) for 24 hours, then I changed the medium to E8 medium. Geltrex was freshly prepared and still usable. The cells were looking very good the day after splitting, and there were a low amount of dead cells.

Two days after splitting, the cells detached completely, and I really do not know what the problem is.

Any suggestions or solutions for this problem? because this problem happened to me 2 times before.

Thanks in advance!

Best,

Aya