maojia

Hi everyone,

I am having problem on culture human iPSCs. My cells are undergoing apoptosis.
Cell lines were thawed recently on feeders and are culturing in standard hESC medium with daily added 5ng/ml bFGF.
Colonies appeared in 2 weeks. After passaging cells in a new feeder well, I start to notice that there are many membrane blebbing in  small colonies and lots of floating cells detected every day. Colonies grow slowly. However, I didn't see cell differentiation.

I tried needle cutting and collagenase IV to passage cells. However, i still can see the blebbing in colonies.
I may try ROCK inhibitor later to see if it will help.

I was wondering if anyone has experience on dealing with iPSCs apoptosis and may know how to save these cells?

Thanks a million
Maojia