Hi,

It's not CHIR99021's fault. Your iPSC cells are very dense. In this situation always, a lot of individual cells begin to swim. A passage is needed (divide 1 well (on photo) into two 6-well plates).

Justyna Augustyniak

I think iPSCs experiencing some death with CHIR99021 and when differentiating in general is pretty normal.

Justyna Augustyniak Hi Justyna,

Thank you very much for your comments. Actually, when iPSC reach 90% confluence, I differentiated them. And after treatment of 6 uM CHIR for 24 hours, 30% of them died (like the image I posted). I tried different seeding density of iPSC, the density of 35000 cells/cm2 will remain 60%-70% alive. But the density of 17000 cells/cm2 will die all.

I am not sure if I should change the concentration of CHIR.

Best,

Longsheng

Stephan Spangenberg Hi Stephan,

Thanks for your views. Actually, I also found it. But I don't know to what extent iPSC will die after treatment of CHIR.

Best,

Longsheng