Tissue processing and scRNAseq
Tumor
TUMOR
Core needle biopsies were obtained from interventional radiology at Massachusetts General Hospital and Brigham and Women’s Hospital and transported in ice-cold hypothermosol before processing. Per patient and time point, the first two to three cores from the operative procedure were allocated for scRNAseq and cut into small pieces with scissors in a 1.5-ml Eppendorf tube containing 1 ml of enzymatic digestion mix (Miltenyi; Human Tumor Dissociation kit). The Eppendorf tubes were then transferred to a rotation shaker set to 37 °C and 550 r.p.m. and shaken for 15 min. The digestion mix was subsequently filtered through a 50-μm Celltrics strainer sitting on a 15-ml falcon tube on ice and mechanically dissociated once more with the plunger of a 1-ml syringe against the screen. The filter and enzymatic mixture were washed with RPMI containing 0.5% bovine serum. The cell suspension was spun at 1,500 r.p.m. (524