Single cell RNA sequencing of primary breast cancer

A total of 9 patients diagnosed with HR+/HER2- breast cancer were recruited for this study. Tissue specimens were obtained by surgical excision without prior treatment. Tumor tissues were mechanically dissociated and enzymatically digested on the day of surgery. We profiled transcriptomic characteristics using single cell RNA-seq to investigate the relationship between migratory tumor subtypes and cancer-associated fibroblasts (CAFs) in tumor progression. Library preparation was performed using the Chromium Single cell 3’ gene expression system (version 2) and processed following the manufacturer’s protocol (10x Genomics, USA). The library was sequenced on a HiSeq 2500 system using 100-bp paired-end sequencing, and raw sequencing data were processed using Cell Ranger.