GSM8422442: HPT5, Pt2_R, scRNAseq, TCR; Homo sapiens; RNA-Seq
All tumors were dissociated using the Human Tumor Dissociation kit (Miltenyi Biotec; Cat#130-095-929) within 30 minutes after surgery. Briefly, minced fragments were incubated with DMEM, enzyme H, enzyme R, and enzyme A and vortexed for 15 minutes at 400 rpm, 37°C. Tissue was filtered (50µm filter, Sysmex; Cat# 04-004-2327), washed, minced with a 1mL syringe plunger, and re-washed. Dissociated cells were then filtered and spun again (30µm filter, Sysmex; Cat# 04-004-2326) at 1500 rpm at 4°C for 5 minutes. Next, incubation with ACK buffer (Gibco, cat#A1049201) was performed followed by removal of the ACK buffer. In all cases, the viability of dissociated single-cell suspensions was increased to >=90% by using a modified protocol of the EasySep Dead Cell Removal (Annexin V) kit (STEMCELL; cat#17899). Gene expression (GEX) and TCR libraries were generated using the 10x Genomics Chromium NextGEM Single Cell V(D)J Reagents kits v1.1 (PN-1000165; PN-1000020; PN-1000005; PN-1000120) according to the manufacturer instructions. Gene expression (GEX) and TCR libraries were generated using the 10x Genomics Chromium NextGEM Single Cell V(D)J Reagents kits v1.1 (PN-1000165; PN-1000020; PN-1000005; PN-1000120) according to the manufacturer instructions.