GSM8829045: Lower hindlimb muscle, injured, ablated_1; Mus musculus; RNA-Seq
Harvested muscles were minced mechanically with scissors and then digested in a 760 U/mL Collagenase Type II enzyme (Worthington Biochemica) solution at 37°C in a shaking water bath for one hour. Wash media (Ham's F10 (Cytiva) + 10% horse serum (Thermo Scientific)) was used to neutralize the digest before resuspending the samples in a second enzyme solution containing Collagenase Type II (Worthington Biochemica) and Dispase (Thermo Scientific) at 37°C in a shaking water bath for 30 minutes. Wash media was again used to neutralize the enzymes and dilute the samples before triturating with a 21G needle via syringe several times. These samples were then passed through a 40um filter to remove larger debris. Cell viability was assessed with propidium iodide, and negative cells were subjected to further processing. Libraries were constructed by the Stanford Universtiy School of Medicine Genomics Core using the 10X Cromium standard protocol, version 2.