GSM8031350
The tumoral biopsies and non-tumoral pleura tissues were minced mechanically into small pieces of 1 cm2 using sterile scalpels in sterile conditions. Further tissue dissociation into single-cell suspension was performed using Multi Tissue Dissociation Kit 2 (Cat No. 130-110-203) combining mechanical with enzymatic dissociation, according to the manufacturer´s datasheet. GentleMACS Dissociators were used for the mechanical dissociation steps. After dissociation, each sample was applied to a filter supplied in the kit to remove any remaining larger particles from the single-cell suspension Cell suspensions were prepared and vitality was assessed for each individual scRNA-seq experiment. Cells were loaded onto a single-cell chip for GEM generation using the 10x Genomics Chromium Controller (10× Genomics, Pleasanton, CA). 3′mRNA-seq gene expression libraries were constructed using the Chromium Single Cell 3′ Library & Gel Bead Kit v2 (10x Genomics) according to the manufacturer's guidelines.