GSM8703057: PBMC, rejection group, 7d, rep3; Rattus norvegicus; RNA-Seq
For PBMCs: Fresh peripheral blood was diluted 1:1 with PBS and layered onto Ficoll lymphocyte separation solution (Beyotime, China). After centrifugation at 400g for 30 minutes at 18°C-20°C, the buffy coat layer (lymphocytes) was aspirated, washed three times with PBS, and stored.For liver tissues: Transplanted liver tissues were minced and digested with 1 mg/mL collagenase B (Roche, Switzerland) in Hanks' Balanced Salt Solution (Solarbio, China) at 37°C for 20 minutes. The digestion mixture was filtered through a 70 μm strainer and resuspended in PBS with 0.04% BSA (Biosharp, China). Single-cell RNA-seq libraries were prepared using the Chromium Single Cell 5′ Library and Gel Bead & Multiplex Kit (10x Genomics, USA). A total of 10,000 cells per sample were loaded onto a Chromium controller. Libraries were sequenced on an Illumina NextSeq platform to achieve sufficient saturation.