Satellite cells were isolated as previously described [27 (link)] from hindlimb muscles. Cells were labeled using the following antibodies: 1:400 CD31-PE (clone 390; eBiosciences), 1:400 CD45-PE (clone 30-F11; BD Pharmingen), 1:4000 Sca-1-PE-Cy7 (clone D7; BD Pharmingen), 1:500 α7-integrin-APC (Clone R2F2; AbLab). Dead cells were excluded by propidium iodide (PI) staining. Cell sorting was performed using a BD FACSAria II cell sorter (Becton-Dickinson). Sorted cells were immediately reanalyzed to ensure purity. Analyses of flow cytometry data were performed using FACSDiva (BD version 8.0.1).
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Isolation and Characterization of Satellite Cells
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CD31-PE (clone 390;
CD45-PE (clone 30-F11;
Sca-1-PE-Cy7 (clone D7;
BD FACSAria II cell sorter
FACSDiva
Antibodies
Cells
Clone
Flow cytometry
Hindlimb
Integrin
Muscles
Propidium iodide
Satellite cells
Sca 1
Corresponding Organization :
Other organizations : Emory University, University of Lübeck, Max Delbrück Center
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Variable analysis
independent variables
- Antibodies used for cell labeling: CD31-PE, CD45-PE, Sca-1-PE-Cy7, α7-integrin-APC
- Satellite cell purity after cell sorting
- Propidium iodide (PI) staining for excluding dead cells
- Positive control: Not specified
- Negative control: Not specified
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