Author: ETB Date: 8/24/95

CTL Stimulators/Feeders

For B3 and GA4 CTLs

Stimulator Cells:
5 x 106 E.G7 cells irradiated with 20,000 rads
Wash 3X in RP-10 (10 ml each)

Feeder Cells:
1 C57/B6 spleen irradiated at 2000 rads
Wash 2X in HBSS (10 ml)

Pool cells in 25 ml of CTL media and aliquot 1 ml per well on 24 well plate.Spleen Cell Preparation
1. Aliquot 5 ml of HBSS to 3 small petri dishes.
2. Wash spleen 1X in first dish.
3. Place spleen on screen in second dish.
4. Grind spleen with plunger of 3cc syringe.
5. Wash second dish with HBSS from third dish.
6. Wash 2X in HBSS (10 ml each).