I am trying to model ex vivo TIL expansion protocol using tumor fragments from C57BL6 bearing ortothopic ID8-p53-/- ovarian tumor.

Briefly, we culture fragments on T cell media (RPMI glutamax, 10% AB-HuS, P/S, Anfot. B - gentamicine, HEPES) suplemented with 6000IU/ml of rhIL-2 (which seemed to work in HM Knochelmann; OncoImmunology, 2021. DOI: 10.1080/2162402X.2021.1959101).

We have observed the initial expansion of lymphocytes, but after the first days in culture,TILs begin to swell and end up dying during the first week.

Any specific recommendation on the media, IL-2 dose or any other component?