Preparation of High Serum for A1 cells: Difference between revisions
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==== For 400 ml ==== | ==== For 400 ml ==== | ||
FCS 40 ml | * FCS 40 ml | ||
* Eagle’s MEM 252 ml | |||
Eagle’s MEM 252 ml | * DdH2O 100 ml | ||
* Pen/ Strep 4 ml | |||
DdH2O 100 ml | * Glutamin 4 ml | ||
* Insulin (stock: 1mg/ ml) 4 ml | |||
Pen/ Strep 4 ml | |||
Glutamin 4 ml | |||
Insulin (stock: 1mg/ ml) 4 ml | |||
* grow at 25ºC in 2% CO2 | * grow at 25ºC in 2% CO2 | ||
Revision as of 10:19, 13 February 2015
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
- 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