GSM7596459: Young, 7 days post-injury (rep2); Mus musculus; RNA-Seq
TA muscles were dissected and placed into 400U/mL collagenase at 37°C for 1h with shaking and then placed into Ham's F-12C supplemented with 15% horse serum to inactivate the collagenase. Cells were passed through three strainers of 100µm, 70µm, and 40µm (BD Falcon) and flow through was centrifuged at 1500×g for 5 min and the cell pellets were re-suspended in Ham's F-12C. To remove dead cells and debris, cells were passed over the Miltenyi, dead cell removal kit columns (Cat# 130-090-101). To remove red blood cells (RBC), cells were incubated with antiTer119 micro magnetic beads and passed over a Miltenyi column (Cat#130-049-901). For the adult and aged uninjured TAs, 6 TA muscles (from 3 mice) were pooled together. For the injured TA muscles 2 TA muscles from 2 different mice were pooled together. Cells were then counted using a BioRad TC20 automated cell counter and processed with the 10X genomics single cell sequencing kit. To capture, label, and generate transcriptome libraries of individual cells we used the 10X genomics Chromium Single Cell 3' Library and Gel Bead Kit v2 (Cat #PN-120237) following the manufactures protocols. single-cell RNA-seq, paired end, Illumina Nextseq500