scRNA-seq
tumor, Tumor
TUMOR, TUMOR
This analysis was performed on one patient with complete response. Fresh tumor tissue was collected on the day of surgery for this patient. Tumor was mechanically and enzymatically dissociated using a human tumor dissociation kit, according to the manufacturer’s instructions (130-095-929, Miltenyi Biotec). In brief, tumor was cut into small pieces and transferred into gentleMACS C tubes containing the enzyme mix. The dissociation was performed using the gentleMACS Octo Dissociator with heaters and with the human tumor dissociation 37C_h_TDK_1 program. Samples were homogenized before being applied to a MACS SmartStrainer 70 µM (130-110-916, Miltenyi Biotec) and placed in a 50-ml tube. Filters were washed with 20 ml of serum-free RPMI (L0500-500, Dutscher) and then centrifuged at 300Library preparation was performed using library prep for 5′ mRNA and VDJ (10x Genomics). Sequencing was performed on an Illumina HiSeq 4000 device. Libraries were sequenced with 100-bp paired-end reads.