help wanted for L6 cell differentiation

[Grad Student Lab 1]

Hi, there. Currently I have a big problem in inducing differentiation and 

fusion of the L6 rat skeletal muscle cells regardless a number of culture 

conditions I have used. The growth medium I used was DMEM containing either 

10% or 20% FBS. I switched the medium to a differentiation medium when the 

cells reached a confluence of 50% or 80% or even 100%. The differentiation 

medium I used was: (1) DMEM + 1% FBS, (2) MEM + 10 horse serum, or (3) MEM + 

4% dialized horse serum. No matte what kind of culture condition and what kind 

of combination I used, all the results were the same: before day 5, the cells 

were in spindle shaped; but starting from day 5, the cells in the centre of 

colonies became round or granular shaped and finally those cells became 

irregular granular shaped while the cells at the edge of colonies remained in

spindle shape and few of them (-20%) got fused. I never can get more than 20% 

of fusion.

I really appreciate if someone out there can give me some thoughts and hints 

in doing this. Many thanks.


p.s.  Please reply to this account or my internet addr.: xzhou at unixg.ubc.ca