GSM5284616: WT-CKD; Mus musculus; RNA-Seq
Kidneys were collected from each experimental animal and weighed, then dissected into small pieces using a razor blade. The dissected tissue was incubated in 1.48 U/mL Liberase DL enzyme mixture (Roche) and 200 U/mL DNase I (Roche) at 37°C for 45 minutes on a rocker. Cell suspension was filtered through 40 μm filters then pelleted by centrifugation at 500 × g for 10 minutes. Cells were resuspended in ACK Lysing Buffer (ThermoFischer) for 3 minutes at room temperature, followed by subsequently wash in DMEM-2% FBS. Cell debris was removed using the Miltenyi Debris Removal Solution. Live and dead cells were counted by Trypan Blue staining (Gibco) using a TC20 Cell Counter (Bio-Rad). Cells were diluted to a concentration of 10^6 cells/mL. We then proceeded to single cell library preparation. The scRNA-seq library and sequencing were performed at the Yale Center for Genome Analysis (YCGA).