GSM9006031: Kidney, CLP 24 hrs; Mus musculus; RNA-Seq
Mice were perfused with ice-cold PBS via cardiac puncture after which the kidneys were harvested. Kidney digest was performed with liberase TM (0.3 mg/mL, Roche, Basel, Switzerland, #291963), hyaluronidase (10mg/mL, Sigma, H4272), DNaseI (20mg/mL) at 37 ̊C for 20 min. After centrifugation, cell pellets were incubated with ACK lysing buffer (ThermoFisher #A1049201) and again centrifuged. Pellets were incubated with 0.25%trypsin EDTA at 37 ̊C for 10 min with shaking at 100 rpm in bacterial shaker. Trypsin was inactivated using 10% fetal bovine serum in PBS. Cells were then washed and resuspended in DPBS supplemented with 0.04% bovine serum albumin. After filtration through a 30 m strainer, cells were loaded onto 10X Chromium System (10X Genomics) at 1000 cells/mcl Library was performed according to manufacturer's instructions (single cell 3' v2 protocol, 10x Genomics)