Protocol for characterizing non-genetic heterogeneity and expression dynamics of surface proteins in mouse muscle stem cells using flow cytometry

Here, we present a protocol for investigating the non-genetic heterogeneity of membrane proteins expression within murine Muscle Stem Cell (MuSC) population isolated from injured skeletal muscles. We describe a method that employs flow cytometry technology to detect variations in membrane CRIPTO protein levels and ensuring measurements standardization. We detail steps for muscle digestion, bulk muscle cell staining, and phenotypic analysis. This approach allows for the identification of MuSC fractions with distinct phenotypic and functional properties.