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Updated: Aug 28, 2025
Identification of Skeletal Muscle Satellite Cells by Immunofluorescence with Pax7 and Laminin Antibodies
Published on: April 19, 2018
Optimized immunofluorescence staining protocol for identifying resident mesenchymal stem cells in bone using LacZ
Chen-Xi Zheng1, Ji Chen2, Jiong-Yi Tian2
1State Key Laboratory of Military Stomatology& National Clinical Research Center for Oral Diseases & Shaanxi International Joint Research Center for Oral Diseases, Center for Tissue Engineering, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Glioma-associated oncogene homolog 1 (Gli1) identifies mesenchymal stem cells (MSCs) in bone. This study optimized immunofluorescence staining to detect these Gli1+ MSCs in mouse bone sections for better in vivo characterization.
Area of Science:
- Stem Cell Biology
- Histology and Tissue Engineering
- Molecular Biology
Background:
- Glioma-associated oncogene homolog 1 (Gli1) is a marker for a specific subpopulation of endogenous mesenchymal stem cells (MSCs).
- These Gli1+ MSCs are characterized by their perivascular location within tissues.
- Understanding the identity and niche of endogenous MSCs is crucial for regenerative medicine and developmental biology.
Purpose of the Study:
- To present an optimized immunofluorescence staining protocol.
- To enable the identification and characterization of resident Gli1+ MSCs in fixed/frozen bone sections.
- To facilitate the in vivo study of endogenous MSCs in LacZ transgenic mouse models.
Main Methods:
- Preparation of fixed/frozen bone tissue sections from LacZ transgenic mice.
- Application of an optimized immunofluorescence staining protocol.
- Utilizing LacZ immunofluorescent staining for cell identification.
Main Results:
- Successful identification of resident Gli1+ MSCs in bone sections.
- Demonstration of the protocol's efficacy in characterizing endogenous MSC identity and niches.
- Validation of the protocol's applicability to diverse organs with LacZ-expressing cells.
Conclusions:
- The optimized immunofluorescence protocol effectively identifies Gli1+ MSCs in mouse bone.
- This method allows for detailed in vivo characterization of endogenous MSC populations.
- The protocol is adaptable for studying LacZ-expressing cells in various organs.
