GSM8819040: AD14CWT-3; Mus musculus; RNA-Seq
For single-cell RNA sequencing (scRNA-seq), kidney tissue from the adenine-induced injury model was collected two weeks post-injury and digested with Liberase, following a specific protocol. The kidney tissue was cut into 1mm pieces on ice, washed with cold PBS. We used dissociation enzyme 0.25% Trypsin (Thermo Fisher, Cat. no.25200-072) and 10 ug/mL lDNase I (Sigma, Cat. no.11284932001) dissolved in PBS with 5% Fetal Bovine Serum (FBS; Thermo Fisher, Cat. no. SV30087.02) to digest the tissues. Kidney tissues were dissociated at 37 ℃ with a shaking speed of 50 r.p.m for about 45 min. Cell suspensions were filtered using a 40um nylon cell strainer and red blood cells were removed by 1X Red Blood Cell Lysis Solution (Thermo Fisher, Cat. no. 00-4333-57). Dissociated cells were washed with 1x DPBS containing 2% FBS. Cells were stained with 0.4% Trypan blue (Thermo Fisher, Cat. no.14190144) to check the viability. Libraries were created using the Chromium Single Cell 3' Library Construction kit (10x Genomics) and sequenced on the NovaSeq6000 (Illumina) platform.