GSM8058097: Health2; Mus musculus; RNA-Seq
Cell populations were isolated from the testes of mice in the ND and HFD groups. In brief, the albuginea was removed, and the tissue was incubated in dissociation buffer containing collagenase IV (Sigma-Aldrich, St Louis, MO, USA; 1 mg/mL), dispase II (10 mg/mL), and DNase I (DN25, Sigma; 50,000 U/mL) for 30 min at 37°C. Enzymatic digestion was quenched with Dulbecco's Modified Eagle's Medium (Gibco, Carlsbad, CA, USA) supplemented with 10% fetal calf serum. Before loading, the cell viability of each sample was evaluated by staining an aliquot with Trypan Blue, and the viability was typically determined to be greater than 80%. The cells were subsequently loaded into each channel of a Chromium Single Cell B Chip (10x Genomics, Pleasanton, CA, USA), and approximately 13,000 cells were recovered from each channel. A Chromium Single Cell 3ʹ Library and Gel Bead Kit using v3 Chemistry (10X Genomics) was used for single-cell barcoding, synthesis of complementary DNA, and library preparation following the manufacturer's protocol (Chromium Single Cell 3ʹ Reagent Kits v3 User Guide, Revision B). The libraries were sequenced on an Illumina NovaSeq 6000 system using paired-end sequencing runs.