GSM8842373: skeletal muscle cells from Clodronate liposome (CLL)-treated mice, biological replicate 3; Mus musculus; RNA-Seq
Freshly isolated quadriceps were minced into small pieces and enzymatically dissociated with 1.5mg/mL Dispase (Gibco, 17105-041) and 0.12% (w/v) Collagenase type I (Sigma-Aldrich, C0130) in two cycles of 30min at 37°C under constant 240 rpm agitation. The suspension was then passed through a 16G needle, followed by a 26G needle and filtered through a 40μm sterile strainer. Then cold CMF was added to inactivate enzymes and red blood cells lysed. To remove cell debris from the suspensions, Ficoll separation was performed. Cells were resuspended in 7 mL of pre-warmed DMEM High Glucose (Gibco, 41965-039) supplemented with 10% FBS (Gibco, 10270-106), in a 15 mL falcon, and 3.5 mL of Ficoll slowly layered on top. Samples were spined at 300g for 30min with no brake nor acceleration and the cell layer formed aspirated to another falcon and washed with cell medium. Chromium Single Cell 3′ Reagent Kit v3.1 (10xGenomics)