Lymphocyte Isolation From Liver and Spleen

Livers were perfused via the portal vein with RPMI 1640 (Thermo Fisher Scientific, Mississauga, ON, Canada) and removed. The livers and spleens were finely minced in RPMI 1640, passed through a 100-gauge steel mesh, and centrifuged at 400g for 5 minutes at 4°C. Cells contained in the supernatant were washed 3 times with RPMI 1640/5% fetal calf serum before being centrifuged on a Percoll (GE Healthcare Canada, Mississauga, ON, Canada) gradient to purify lymphocytes.¹⁰