GSM8995897: 20m_4; Rattus norvegicus; RNA-Seq
Rats were euthanized by CO₂ inhalation, and tumors were harvested as previously described. Briefly, tumors were enzymatically digested in DMEM/F12 medium containing 2 mg/mL collagenase IV, 2 mg/mL hyaluronidase, and 2 mg/mL bovine serum albumin (BSA) at 37ºC for 1–2 hours, followed by freezing in cryopreservation medium (90% fetal bovine serum (FBS) and 10% dimethyl sulfoxide) in liquid nitrogen. For mammary tumor scRNA-seq, viable cells were resuspended in PBS with 0.04% BSA at a cell concentration of 1000 cells/μL. 10,000 cells were loaded onto a 10x Genomics Chromium™ instrument (10x Genomics) according to the manufacturer's instructions. The scRNA-seq libraries were prepared according to the Chromium Next GEM Single Cell 5' HT v2 protocol (10x Genomics). Quality control for amplified cDNA libraries and final sequencing libraries were performed using Bioanalyzer High Sensitivity DNA Kit (Agilent). The pooled libraries were sequenced on Illumina NovaSeq X plus platform.