[Cell-biology] Primary mouse myoblast culture

[imperat from hotmail.com]

Hi all,

I'm in the process of growing primary myoblasts isolated from lower
limb muscles, and repeatedly run into problems. Specifically, after a
couple of months of successful work, it becomes increasingly difficult
to maintain a line in any particular incubator, such that even freshly
isolated myoblasts tend to gradually die. Is there any precedence and/
or solution to this problem? I am growing them, on Matrigel-coated
plates, under the following conditions:
   -- 20% FBS
   -- 10% horse serum (HyClone)
   -- hi-glucose DMEM
   -- 10ng/ml bFGF
   -- 5% CO2

Thanks in advance,
Don