Primary mouse myoblast culture

[imp...@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