Fresh tissues were immediately used to prepare their single cell suspension. Briefly, the collected fresh tissue was washed by DPBS (PBS without calcium and magnesium) and carefully chopped into small pieces. Enzymatic digestion was performed with collagenase I (2 mg/mL), collagenase II (2 mg/mL), collagenase IV (2 mg/mL) and dispase (0.2 mg/mL) under 37 °C for 15-25 minutes. The digested solution was transferred to a centrifuge tube through a 40 μm stainless nylon mesh (Greiner Bio-OneGmbH, Germany). The filtrate was immediately centrifuged at 500 g for 5 min, and discarded the supernatant slowly. The cell sediment was resuspended and lysis with RBC Lysis buffer on ice for 5 min to remove red blood cells, and then washed with DPBS at 400 g for 5 min twice. Cell concentration and viability were determined on a Cellometer Auto 2000 (nexcelom, USA) after AO/PI staining. Immediately after dissociation, cells were resuspended in FACS buffer (5% FBS in PBS) and Fc-blocked for 10 min. This was followed by a wash and staining with anti-CD45 PE (BioLegend; San Diego, CA) for 30 min in sorting buffer (0.1% BSA in PBS) at 4 °C. After washing, viable CD45+  cells were sorted using the BDFACSAria Ⅲ. Subsequently, the cells were washed twice and re-suspended in a sorting buffer. A cell number and viability count were performed on a Cellometer Auto 2000 using the ViaStain™ AOPI Staining Solution (Nexcelom Bioscience LLC, Lawrence, MA, USA) immediately prior to scRNAseq. Library was performed according to the manufacter's instructions (single cell 5' v2 protocol, 10x Genomics). Briefly, single-cell suspensions were resuspended in the master mix andloaded together with partitioning oil and gel beads into the chip to generate the gelbead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in everysingle GEM was retrotranscripted to cDNA, which contains an llumina R1 primersequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooledbarcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR andthe apropiated sized fragments were selected with SPRIselect reagent forsubsequent library construction. During the library construction llumina R2 primersequence, paired-end constructs with P5 and P7 sequences and a sample indexwere added.