Preparation of High Serum for A1 cells: Difference between revisions
Jump to navigation
Jump to search
No edit summary |
No edit summary |
||
| Line 8: | Line 8: | ||
* Insulin (stock: 1mg/ ml) 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) | * wash with APBS (100 ml PBS + 25 ml ddH2O) | ||
Revision as of 10:20, 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