I've kept them for up to two weeks. Not sure how long they can go for. Though it's probably somewhere in the ballpark. Test it, and replace media ever 2-3 days or so.

Thank you Martin!

There's several different protocols to differentiate them too. What have you used - which protocol? And the two week duration that you've kept them is AFTER they're differentiated?

Thanks so much!

Reeteka

10uM of RA for 2-4 days. media is always the same 1:1 EMEM:F12 + 10% FBS + 1% antibiotic.

Yes, I've kept them two weeks after differentiation. May go longer, but you'd have to test it, Why do you need to culture them that long? Are you looking to do electrophysiological work?

Dear Martin

do you add BDNF as final "touch" for differentiation?

And do you differentiate them in 96 well plate? AT what initial density?

Thanks

I haven't used BDNF yet, though I hear it helps if you administer it in a sequential fashion. Say 2 days of RA, then 2 days of BDNF. For a 96-well, I'd say a good starting density is 5K.