title: GSM8016963: Vascular，rep2; Mus musculus; RNA-Seq
library_construction_protocol: To obtain single cells from an aorta, the whole aortas were minced into small pieces and then digested in 1X Aorta Dissociation Enzyme Stock Solution at 37°C for 1 h. Cell suspensions were prepared by filtering them through a cell strainer. To obtain single cells from a spleen, the whole tissue was ground and cell suspensions were prepared by filtering them through the same cell strainer as mentioned before. Then, cell suspensions were added to the lymphocyte separation medium and mononuclear cells were obtained after density gradient centrifugation. Cell capture and cDNA synthesis were performed using a single-cell 5' Library and Gel Bead Kit (10× Genomics, 1000006) and Chromium Single Cell A Chip Kit (10× Genomics, 120236). The cell suspension (300-600 living cells per microliter determined by CountStar) was loaded onto the Chromium single-cell controller (10× Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol. In short, single cells were suspended in PBS containing 0.04% BSA. Approximately 10,000 cells were added to each channel, and the target cells recovered were estimated to be approximately 6,000 cells. Captured cells were lysed, and the released RNA was barcoded through reverse transcription in individual GEMs.