UMR cell culture medium
5% MEM
12.5 ml 1 M HEPES
50 ml FBS
10 ml P/S
10 ml NEAA
Meium (Minimum essential medium eagle) to 1000 ml
Frozen medium
5% DMSO 0.5 ml
5% glycerol 0.5 ml
10% FBS 1 ml
80% 0% MEM 8 ml
Or Marika’s frozen medium:
5% MEM 2 ml
FBS 7 ml
DMSO 1 ml
filter
To freeze cells from 175 ml flask
Trypsinize cells (7 ml Trypsin + 5 % MEM).
spin 1200 rpm 4 min
resuspend cells in freezing medium 5 ml.
1 ml / vial
use paper to wrap the vials and put in the middle of a box full of bulbs. -70C overnight. Transfer the vials into liquid nitrogen.
To thaw cells:
Immediately warm the vial at 37C water bath with hand shaking. As soon as the soln melt, wipe the vial with ethanol, transfer the soln in a flask containing MEM. The next day change the medium.
01001 CHAPTER 26 PAGE 4 01001 DEPARTMENT OF AGRICULTURE
01001 CHAPTER 335 PAGE 15 01001 DEPARTMENT OF AGRICULTURE
01001 CHAPTER 339 PAGE 15 01001 DEPARTMENT OF AGRICULTURE
Tags: culture medium, medium, culture