View source for Preparation of High Serum for A1 cells

== Material == 
''For 400 ml''
* FCS 				40 ml
* Eagle’s MEM			252 ml
* DdH2O				100 ml
* Pen/ Strep			4 ml
* Glutamin			4 ml
* Insulin (stock: 1mg/ ml)	4 ml

== Procedure ==
# grow at 25ºC in 2% CO2
# if you want to grow without CO2, then substitute L-15 for the MEM

==== passage once per week 1:3 ====

# wash with APBS (100 ml PBS + 25 ml ddH2O)
# trypsinize with TE
# stop with media
# centrifuge cells at 100 rpm 3 minutes

==== to make myotubes: ====

# wash cells 1x with APBS
# put cells in 0.5% FCSmedia
# wait 3 days