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Tumor organoid-T-cell coculture systems.
Nature Protocols ( IF 10.419 ) Pub Date : 2019-12-18 , DOI: 10.1038/s41596-019-0232-9
Chiara M Cattaneo,Krijn K Dijkstra,Lorenzo F Fanchi,Sander Kelderman,Sovann Kaing,Nienke van Rooij,Stieneke van den Brink,Ton N Schumacher,Emile E Voest

T cells are key players in cancer immunotherapy, but strategies to expand tumor-reactive cells and study their interactions with tumor cells at the level of an individual patient are limited. Here we describe the generation and functional assessment of tumor-reactive T cells based on cocultures of tumor organoids and autologous peripheral blood lymphocytes. The procedure consists of an initial coculture of 2 weeks, in which tumor-reactive T cells are first expanded in the presence of (IFNγ-stimulated) autologous tumor cells. Subsequently, T cells are evaluated for their capacity to carry out effector functions (IFNγ secretion and degranulation) after recognition of tumor cells, and their capacity to kill tumor organoids. This strategy is unique in its use of peripheral blood as a source of tumor-reactive T cells in an antigen-agnostic manner. In 2 weeks, tumor-reactive CD8+ T-cell populations can be obtained from ~33-50% of samples from patients with non-small-cell lung cancer (NSCLC) and microsatellite-instable colorectal cancer (CRC). This enables the establishment of ex vivo test systems for T-cell-based immunotherapy at the level of the individual patient.
更新日期:2019-12-19

 

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