How cancer evades immunesystem detection and spreads NatureComms
. TIL-CM comprises RPMI 1640 with GlutaMAX supplemented with 2 mM HEPES, 10ug/ml gentamicin and 1% Antibiotic-Antimycotic and 10% human AB serum . After 5 days of initiation, half the media was removed and refreshed with TIL-CM and IL-2. Subsequently, half the media was exchanged every 2–4 days. After 14-18 days, 1 to 2 million TILs were transferred to T75 flasks containing irradiated allogenic PBMC feeders at a ratio 1 to 200 for the Rapid Expansion Protocol .
For clustering of all cell types, Seurat alignment across patients was applied. The Seurat object with all the cells was first split by patient ID, then for each patient’s object, raw UMI counts were lognormalized and variable genes were called independently based on average expression > 0.1 and average dispersion >1.
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