I am performing hematopoietic differentiation of hESCs using OP9 coculture system and I am having a big problem. The cells start to detach at day 3-4 of coculture, first the colonies and finally the whole stroma detach. I'm desperate because I'm trying to figure out what it is all about. I have OP9 with differentiation medium for 10 days and they look totally right, the hESCs are growing almost at the normal rate.
I will be so pleased if anyone can give me a hand, I really don't know what's going on. Thanks in advance!