Primary muscle cultures - Keeping myoblasts alive! (Oct/26/2009 )

Hello everyone

I'm having a bit of trouble establishing primary muscle cultures and wondered if anyone could help.

Basically, I've developed a protocol for isolating the hind limb muscles from neonate mice, enzymatically digesting them and plating them out. I then have trouble keeping them at the right confluency - too dense and they differentiate, too sparse and become senescent!!! It seems to be an incredibly delicate balance.

Does anyone have any muscle/myoblast culturing experience - I could do with some hints and tips on how to keep them growing!

BW
Emily

This is an old post so I don't know if you're still watching it, but... I haven't had as much trouble keeping them at the right density. Could you expand a little on the details of your protocol? They don't seem to become senescent for me until a few passages in, even if plated at somewhat low densities (obviously not too low). There are so many different protocols though for isolating primary myoblasts so let me know the details first.