GSM7091135: Bl6, sham, replicate 2, scRNAseq; Mus musculus; RNA-Seq
Skeletal muscle tissue was processed to generate single-cell suspensions using mechanical dissociation and enzymatic digestion. Live cells (PI- cells) were isolated by fluorescence-activated cell sorting using a SonySorter SH800s. For mouse HLI experiments, an equal number of PI- cells and PI-/CD31-/CD45- cells were pooled for each sample to increase the representation of relatively scarce non-hematopoietic and non-endothelial cell-types of interest (e.g. muscle stem cells and fibro-adipogenic progenitors). A total of approximately 150,000 cells were sorted for each sample, precisely counted using a hemocytometer, and used for scRNA-seq analysis. Single-cell RNA sequencing libraries were generated using Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10x Genomics) according to the manufacturer's protocol. Approximately 16,000 - 24,000 cells per sample were used for GEM creation.