GSM9008269: SARS-CoV-2 infection, Meplazumab and 3E8 treatment-2 [scRNA-seq]; Macaca mulatta; RNA-Seq
The scRNA-seq was performed by Genedenovo Biotechnology Co., Ltd. The lung tissues of rhesus macaques (Healthy, n=4; Ctrl, n=4; MPZ, n=4; 3E8, n=4; MPZ+3E8, n=4; at 7dpi) and virus-infected hCD147 mice (n=4; at 3dpi) were cut into 1–2 mm3 pieces in DPBS buffer and digested with collagenase I and DNase I at 37°C for 30–60 min with persistent shaking every 10 min. The digested tissues were then passed through a 70 μm mesh filter, and the cell suspension was centrifuged at 4 °C for 5 min (350 g). The cell pellet was suspended in red blood cell removal solution at 4 °C for 10 min and then resuspended in DPBS buffer (0.04% BSA and 0.01 mM EDTA) after being washed by DPBS buffer. Following the depletion of the dead cells, the cell suspensions were directly performed for scRNA-seq according to the manufacturer's instructions. For the samples from rhesus macaques, the cells in single-cell suspension were counted by trypan blue dye at a proper concentration of 1000–2000 cells/μL. Single-cell suspensions were loaded onto 10× Genomics Chromium v3.1 system to generate single-cell Gel Beads-In-Emulsions (GEMs), where all generated full-length cDNA share a common 10× barcode. Then, GEMs were disrupted and cDNA was amplified via PCR. The quality control of the sequencing libraries was performed by High Sensitivity DNA assay Kit (5067-4626, Agilent Technologies), and the concentration quantification was conducted using ABI StepOnePlus Real-Time PCR System. Finally, the sequencing was performed on Illumina sequencing platform in PE150 mode. For the samples from hCD147 mice, cell suspensions were barcoded through the SeekOne single cell platform.