title: GSM8167386: Testis_1Month_scRNA; Mus musculus; RNA-Seq
library_construction_protocol: Testis samples from C57BL/6j mice were dissected aseptically using sterilized scissors. The samples were washed three times with DPBS (Thermo Fisher Scientific, 14190-144), and the tunica albuginea was carefully removed using sterilized tweezers to isolate the interstitial cells as our previous reported13, 14. The interstitial cells were then digested with 1 mg/ml collagenase type IV (Thermo Fisher Scientific, 17104019) in DMEM-F12 (phenol red-free) (Thermo Fisher Scientific, 21041025) at 37 °C for 10 minutes. During the digestion process, the tissues were gently pipetted up and down. The digestion process was halted by adding 2% FBS, and the resulting cell suspension was filtered through a 40-μm nylon mesh. The cells were then centrifuged at 350 rcf for 5 minutes at room temperature and suspended in DPBS or 2% FBS in DPBS for subsequent fluorescence-activated cell sorting analysis (FACSAria III, BD Bioscience). For the 10x Genomics platform, the cell count for each sample was determined using the Biorad Automated Cell Counter. After GEM generation and barcoding utilizing the Chromium Single Cell Library & Gel Bead Kit v3.1 (10x Genomics) according to User Guide, approximately 20,000 single cells per sample were captured by Chromium Controller (10x Genomics). For library construction, utilizing the Chromium Single Cell Library & Gel Bead Kit v3.1 (10x Genomics) according to User Guide.