GSM8898886: db/db_scRNA-seq_replicate 2; Mus musculus; RNA-Seq
We pooled aortas from 8 mice into two replicates (4 mice per replicate) for each of the three groups. Aortas (4/group) were cut into ~1 mm pieces and placed into enzyme mixture containing Liberase DH (1Unit/aorta, 5401054001, Roche), Elastase (0.125 mg/aorta, E0127, Sigma) and DNase I (Kunitz unit/aorta, 79254, Qiagen) in M199 medium (10060CV, Corning). After incubating at 37°C for 45 minutes, cell suspensions were filtered through a 70-µm strainer and centrifuged at 500g for 5 min. The cell pellets were washed with Hank's balanced salt solution (HBSS) and FACS buffer (HBSS containing 2% FBS) and resuspended in 500 µL of FACS buffer and passed through 35-µm strainer. Cell viability was measured using the Acridine Orange/Propidium Iodide method (Cellometer, Nexcelom Biosciences). Around 10,000 single cells were used for library preparation using the Chromium Next GEM single cell 3' reagent kits v3.1 (Dual index), Version CG000315 Rev B (10X Genomics) following the manufacturer's protocol.
GEO
GSM8898886_r1