TIL isolation.

Tumors were excised immediately after mice were euthanized and placed into CTL media. They were diced into small pieces, incubated with digestion media (DMEM, 25 mg/l Hyaluronidase, 1 g/l Collagenase IV; all from Life Technologies), incubated at 37°C for 30 minutes with shaking, and then centrifuged for 10 minutes at 500 g. Tumor samples were washed once with RPMI (Life Technologies), filtered through a 70-μm filter (Falcon), and plated for 1 hour at 37°C, 5% CO₂ to remove adherent tumor cells.