GSM5692602: Kidney, CTRL; Mus musculus; RNA-Seq
Injured kidneys were harvested, minced into approximately 1 mm3 cubes, and digested using Liberase™ (100 µg/mL) and DNase I (10 µg/mL) (Roche Diagnostics) for 25 min at 37 °C. Reaction was deactivated by adding chilled DMEM with 10% FBS. The solution was then passed through a 40-µm cell strainer. After centrifugation at 1,000 RPM for 10 min at 4 °C, the cell pellet was resuspended in chilled DMEM with 10% FBS and passed through another 40-µm cell strainer. Cell number and viability were analyzed using trypan blue staining (Invitrogen). This method generated single cell suspension with greater than 75% viability. The scRNA-seq library and sequencing were performed at the Yale Center for Genome Analysis (YCGA).