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

使用流式细胞术表征小鼠肌肉干细胞中非遗传异质性和表面蛋白表达动力学的协议

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作者:Laura Pisapia, Vincenzo Mercadante, Gennaro Andolfi, Gabriella Minchiotti, Ombretta Guardiola

Abstract

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 protocol that employs flow cytometry technology to detect variations in membrane CRIPTO protein levels and ensure 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. For complete details on the use and execution of this protocol, please refer to Guardiola et al.1.

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