Hi Haseeb, thank you for the reply.
The line I’m using is a patient-derived iPSC line that we reprogrammed ourselves here in the lab. I didn’t use the same line that you are using. They have good morphology prior to differentiation (no cratering, no spontaneous differentiating regions) and are kept at 70-80% confluency prior to routine passaging.
I’m unsure if it’s the flow protocol or the differentiation protocol that’s the issue, because I saw beating clusters on the edges of the plate and there was no beating cells in the middle at all. I timed the media changes from Chiron supplemented media to IWP2 media as close to 48 hrs as possible, and I’ve used the flow protocol before to detect cTNT+ cells derived using a commercial differentiation kit and they were able to detect cTNT+ cells.
if it’s not too much trouble, could you share the flow protocol used by Allen Cell to evaluate differentiation efficiency?
Thank you!