Identification and Analysis of Myogenic Progenitors In Vivo During Acute Skeletal Muscle Injury by High-Dimensional Single-Cell Mass Cytometry

Stig Henrik Andersen; Troels R&#248;nn Kj&#230;r; Kedar Ghimire; Ermelinda Porpiglia

doi:10.3791/65944

Identificación y análisis de progenitores miogénicos in vivo durante la lesión aguda del músculo ...

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Identification and Analysis of Myogenic Progenitors In Vivo During Acute Skeletal Muscle Injury by High-Dimensional Single-Cell Mass Cytometry

Identificación y análisis de progenitores miogénicos in vivo durante la lesión aguda del músculo esquelético mediante citometría de masa unicelular de alta dimensión

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11:02 min

December 1, 2023

DOI: 10.3791/65944-v

Stig Henrik Andersen¹, Troels Rønn Kjær¹, Kedar Ghimire¹, Ermelinda Porpiglia^1,2

¹Department of Biomedicine,Aarhus University, ²Aarhus Institute of Advanced Studies,Aarhus University

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Overview

This research investigates the molecular mechanisms of muscle stem and progenitor cell dysfunction in aging and disease. Through the application of single-cell mass cytometry and FACS, the study focuses on how these cells interact within the muscle microenvironment and their roles in regeneration dynamics, providing insights into potential therapeutic interventions.

Key Study Components

Research Area

Muscle biology
Stem cell function
Regeneration dynamics

Background

Stem cell dysfunction impacts tissue regeneration in aging and disease.
Effective cell-cell communication is crucial within the stem cell niche.
Previous technologies lacked single-cell resolution for detailed study.

Methods Used

Single-cell mass cytometry for high-dimensional analysis.
FACS (Fluorescence-activated cell sorting) for cell purification.
In vivo mouse models for regeneration studies.

Main Results

Identification of unique molecular signatures in muscle stem cells.
Insights into the quiescence and reactivation of stem cells.
Characterization of cell state transitions vital for effective regeneration.

Conclusions

This study demonstrates the utility of advanced cytometry techniques in understanding muscle stem cell biology.
Findings are relevant for developing therapeutic strategies for age-related muscle degeneration.

Frequently Asked Questions

What is the main focus of the study?

The study focuses on the molecular mechanisms and interactions of muscle stem and progenitor cells in aging and disease.

How does single-cell mass cytometry contribute to this research?

It allows for high-dimensional analysis of multiple protein parameters in individual cells within complex tissues.

What are the implications of understanding cell state transitions?

Understanding these transitions can reveal targets for interventions to improve tissue regeneration.

What technologies were used in this study?

The study utilized single-cell mass cytometry and FACS for cell purification.

Which biological system was investigated?

The investigation was conducted using skeletal muscle stem and progenitor cells in mouse models.

Why is cell-cell communication important?

It is crucial for the effective regeneration of muscle tissue by maintaining proper stem cell function.

What is a major outcome of the research?

Identification of unique molecular signatures that can be used to study stem cell quiescence and activation.

El protocolo aquí presentado permite la identificación y el análisis de alta dimensión de células madre y progenitoras musculares mediante citometría de masas unicelulares y su purificación por FACS para estudios en profundidad de su función. Este enfoque se puede aplicar para estudiar la dinámica de regeneración en modelos de enfermedad y probar la eficacia de las intervenciones farmacológicas.

Nuestra investigación investiga los mecanismos moleculares que subyacen a la disfunción de las células madre en el envejecimiento y la enfermedad. Nuestro objetivo es comprender cómo las células madre y progenitoras musculares interactúan con otras células en el microambiente muscular para lograr una regeneración efectiva. También estamos estudiando si una comunicación celular estéril afecta la función de las células madre musculares en el envejecimiento y la enfermedad.

Una barrera importante para la investigación de la comunicación entre células dentro del nicho de las células madre ha sido la falta de tecnologías que proporcionen resolución de una sola célula. La citometría de masa de una sola célula permite el análisis cuantitativo simultáneo de alto rendimiento en múltiples tipos de células y fenotipos moleculares en tejidos complejos. Nuestro protocolo permite la purificación y el perfil profundo de las células madre raras y la progenie durante la regeneración muscular utilizando anticuerpos contra los marcadores de la superficie celular y los factores de transcripción miogénicos.

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