The single cell landscape of the human vein after arteriovenous fistula creation and implications for maturation failure.

We performed single-cell RNA sequencing of venous samples from 20 patients, including 6 pre-access veins, 2 early AVF resections, and 12 second-stage AVFs that matured (6) or failed (6). Overall design: Vessels were collected in cold cell culture media and enzymatically digested to generate single-cell suspensions. Samples were run using the Chromium Single Cell 3' Library & Gel Bead Kit v3 (10X Genomics) with a target capture of 10,000 cells. Libraries were sequenced on the Illumina NovaSeq 6000/X Plus. FASTQ files were generated with Cell Ranger's mkfastq pipeline (version 6.0.2). The Cell Ranger's count pipeline (version 6.0.2) was used to generate raw gene-barcode matrices from alignment to the 10X Genomics pre-built Cell Ranger human reference package (version 2020-A), from GRCh38 Ensembl build 98/GENCODE v32 gene annotations.