Author: ETB Date: 9/19/95

Milking Cells

Citrate-phosphate milking buffer:
0.131 M citric acid/0.066 M Na2HPO4
pH ª 3.31. Wash cells 4X in 50 ml HBSS (spin 10 minutes at 3)

2. Resuspend 2 x 108 cells/5 ml of milking buffer for 5 minutes

3. Spin for 3 minutes at 4

4. Remove milk and freeze at - 20∞

5. Quickly resuspend pellet in appropriate media

Protocol taken from W.J. Storkus ET. AL. J Immunother, Vol. 14, No. 2, 1993.