FROZEN  FEEDERS





THAW OUT  CELLS IN THE NORMAL WAY, AFTER CENTRIFUGATION ,  ASPIRATE  

OFF THE SUPERNATANT AND RESUSPEND CELL PELLET IN  120 MLS OF MEDIA.   

THIS SHOULD GIVE YOU A DENSITY  OF:   3.5 X 10 5 CELLS/ML, BECAUSE  

THE TOTAL NUMBER OF CELLS  =  4.2 X 10 7 CELLS.     PLATE OUT THE 

CELLS ON THE  10CM PLATES, 12 MLS/PLATE AND SWIRL  PLATES.    

REMEMBER TO USE PLATES THAT HAVE BEEN PRE-TREATED WITH GELATIN  

(2 HOURS PRIOR THIS).



1 VIAL = 1 ML =   4.2  X 107  CELLS  =  10 X 10 CM PLATES



10  X 10CM PLATES  =  10 X 6-WELLS PLATES  =  10 X  24  

WELL PLATES  =  30 X 6 CM



1 X 10CM PLATE  =  12 MLS  =  1 X 6-WELLS PLATE  =  

1 X 24 WELL PLATE  =   3 X 6 CM



The 96-wells plates and 48-wells plates require a dilution, because 

of different cell density.



Feeders/STO Media:   DMEM;  7%  FCS,  1 X GPS