GSM5025052: BL1, Her1 (CITE-seq); Homo sapiens; OTHER
To enrich immune cells from human pancreatic tissues, pancreatic tissues were enzymically digested as described in the previous study. After removing red blood cells with RBC lysis buffer (Sigma, R7757), the recovered cells were then washed again with RPMI buffer that contained 10% BCS and suspended in the recovery cell culture freezing medium (Gibco, #12648-010) to be stored in -80 ºC freezer and liquid nitrogen. To isolate pure leukocytes for the single-cell sequencing analyses, the frozen single cells were thawed and washed twice with RPMI containing 10% FBS. The recovered cells were resuspended in HBSS containing 2% BCS and stained with Live/Dead marker (BioLegend, Zombie Aqua, #423102) and anti-CD45 antibody (BioLegend, PerCP-Cy5.5, #304028) for FACS. Library generation for feature barcoding, gene expression, and V(D)J TCRs was performed using the 10x Chromium Single Cell reagent kits. Following library generation, sequencing was performed on an Illumina HiSeq 4000 (2 x75bp).