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Reporter-Based Isolation of Developmental Myogenic Progenitors
Eyemen Kheir1,2, Gabriella Cusella3,4, Graziella Messina5
1Centre for Integrative Biology (CIBIO), University of Trento, Trento, Italy.
Frontiers in Physiology
|April 21, 2018
Summary
This protocol details purifying embryonic and fetal skeletal muscle progenitors using fluorescence-activated cell sorting (FACS). This method enables molecular and phenotypic characterization of myogenic progenitors for developmental studies.
Area of Science:
- Developmental biology
- Cell biology
- Biotechnology
Background:
- Mammalian tissue formation requires precise regulation of cell interactions.
- Isolating distinct progenitor and stem cell populations is crucial for studying their properties.
- Skeletal muscle development involves complex cellular processes and progenitor differentiation.
Purpose of the Study:
- To describe a protocol for purifying myogenic progenitors from embryonic and fetal skeletal muscle.
- To enable the molecular and phenotypic characterization of these progenitors.
- To provide a method for investigating developmental myogenic processes.
Main Methods:
- Utilizing fluorescence-activated cell sorting (FACS) for progenitor purification.
- Employing murine strains with fluorescent reporter genes specific to myogenic progenitors.
- Detailed dissection and enzymatic dissociation procedures to maximize single-cell yield.
Main Results:
- Successful isolation of myogenic progenitors based on fluorescent reporter expression.
- Optimization of tissue dissociation for efficient FACS.
- Establishment of a ~6-7 hour protocol for progenitor purification.
Conclusions:
- This FACS-based protocol provides a robust method for isolating developmental myogenic progenitors.
- The purified progenitors can be used for subsequent molecular and phenotypic analyses.
- This technique advances the study of skeletal muscle development and progenitor biology.
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