Single-cell and single-nucleus RNAseq of the human and mouse skeletal muscle across age
Single-cell and single-nucleus RNAseq of the human and mouse skeletal muscle across age
Human intercostal muscle biopsies (inner part between the second and third ribs) were collected with consent from deceased transplant organ donors by the Cambridge Biorepository for Translational Medicine, Cambridge, UK (CBTM), immediately placed in HypoThermosol FRS preservation solution and shipped to Sanger Institute for processing.  Skeletal muscle tissue was processed according to the following protocol for single-cell isolation: https://www.protocols.io/view/single-cell-isolation-from-human-skeletal-muscle-261geyydv479/v1). Briefly, muscle tissue was minced, digested in the solution of Collagenase II (Worthington-Biochem, LS004176) and Dispase (Gibco, 17105041) and, following that, the lysate was centrifuged in the gradient of Percoll to recover a fraction with single-cells. Cell or nuclei suspension were  loaded on the Chromium Controller (10X Genomics) with a targeted cell or nuclei recovery of 8000.  10X Genomics Single Cell 3â€™ v2 or v3 Reagent Kit was used to create GEM, perform cDNA synthesis and generate sequencing libraries according to standard instructions.