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JoVE Journal
Developmental Biology
Aislamiento, expansión y diferenciación de progenitores fibro-adipogénicos a partir del modelo de...
Aislamiento, expansión y diferenciación de progenitores fibro-adipogénicos a partir del modelo de...
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
Fibro-Adipogenic Progenitor Isolation, Expansion, and Differentiation from the Spiny Mouse Model

Aislamiento, expansión y diferenciación de progenitores fibro-adipogénicos a partir del modelo de ratón espinoso

Full Text
3,444 Views
07:00 min
November 15, 2024

DOI: 10.3791/66717-v

Bruce Lin1, Hesham Soliman1,2, Fabio M. V. Rossi1, Marine Theret1

1School of Biomedical Engineering and Department of Medical Genetics,University of British Columbia, 2Aspect Biosystems

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol outlines the isolation of skeletal and cardiac muscle fibro-adipogenic progenitors (FAPs) from spiny mice (Acomys) using enzymatic dissociation and fluorescence-activated cell sorting. The method provides reliable enzyme digestion and antibody staining for effective sorting and in vitro culture of FAPs.

Key Study Components

Area of Science

  • Cell biology
  • Regenerative medicine
  • Stem cell research

Background

  • Understanding cell communication and behavior is crucial for tissue homeostasis.
  • Fibro-adipogenic progenitors play a significant role in tissue repair.
  • The spiny mouse model presents unique challenges due to limited research tools.
  • Reliable protocols are needed for studying FAPs in this model.

Purpose of Study

  • To develop a robust protocol for isolating FAPs from spiny mice.
  • To enable further analysis of FAP differentiation into myofibroblasts and adipocytes.
  • To provide a foundation for studying other cell types in the spiny mouse model.

Methods Used

  • Enzymatic dissociation of muscle tissue.
  • Fluorescence-activated cell sorting (FACS) for FAP isolation.
  • Testing various enzyme combinations and concentrations.
  • Utilization of multiple antibodies for effective sorting.

Main Results

  • Successful isolation and expansion of FAPs from spiny mice.
  • FAPs can be differentiated into myofibroblasts and adipocytes.
  • Establishment of a reliable enzymatic digestion protocol.
  • References for robust antibody staining methods provided.

Conclusions

  • The developed protocol enhances the study of FAPs in spiny mice.
  • Future studies can explore additional markers and cell types.
  • This work contributes to understanding tissue repair mechanisms.

Frequently Asked Questions

What are fibro-adipogenic progenitors?
Fibro-adipogenic progenitors (FAPs) are stem cells that can differentiate into fibroblasts and adipocytes, playing a role in tissue repair.
Why use spiny mice for this research?
Spiny mice are a unique model for studying tissue regeneration due to their remarkable healing capabilities.
What is fluorescence-activated cell sorting?
Fluorescence-activated cell sorting (FACS) is a technique used to sort a mixture of cells into different populations based on specific markers.
How can FAPs be differentiated?
FAPs can be differentiated into myofibroblasts and adipocytes through specific culture conditions and growth factors.
What are the implications of this research?
This research provides insights into tissue repair mechanisms and may lead to advancements in regenerative medicine.

Este protocolo describe el aislamiento de progenitores fibroadipogénicos (FAPs) del músculo esquelético y cardíaco a partir de ratones espinosos (Acomys) mediante disociación enzimática y clasificación celular activada por fluorescencia. Los FAPs obtenidos de este protocolo pueden expandirse y diferenciarse eficazmente a miofibroblastos y adipocitos.

Nuestra visión general es comprender y comprender mejor la comunicación celular y el comportamiento celular que conduce a la metástasis de los tejidos y cómo los procesos celulares están mal regulados en la reparación de la alimentación. Para ello, estamos utilizando varios modelos de los cuales uno es el ratón espinoso. Uno de los principales problemas de este nuevo modelo de ratón es la falta de herramientas para estudiarlo.

Aquí proporcionamos un nuevo protocolo, que contiene una digestión enzimática confiable y tinción de anticuerpos. Lo que nos permite clasificar con éxito los progenitores fibro-adipogénicos y cultivarlos in vitro para un análisis más detallado de su destino. Hemos probado varias combinaciones y concentraciones de enzimas, así como múltiples anticuerpos, y nuestro método proporciona referencias para el protocolo más fiable y robusto.

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