collection of skeletal and cardiac muscles from spiny mouse for isolating fibro-adipogenic progenitors (faps)

<meta charset="utf8"></head><body>Digestion du cœur et des muscles squelettiques de souris pour la culture de progéniteurs fibro-adipogènes

Initially recognized for its exceptionally fragile skin and remarkable ability to repair skin injuries, spiny mouse has demonstrated superior regenerative capacity in various organ systems, such as musculoskeletal, renal, central nervous system, and cardiovascular, when compared to Mus musculus1,2.
Fibro-adipogenic progenitors (FAPs) are a subset of stromal cells that are resident in various tissues, including skeletal and cardiac muscle. These cells possess a unique capacity to commit to fibrogenic and adipogenic lineages in vivo and in vitro3,4. FAPs play a crucial role in tissue regeneration by modulating the extracellular matrix and supporting the functions of other cell types involved in the repair process. In skeletal muscle, FAPs become activated in response to injury and facilitate muscle stem cell differentiation and myogenesis5,6. In ischemic injury to the heart, FAPs lay down scar tissue to maintain the integrity of the myocardium. In contrast to muscle, cardiac FAPs become chronically activated and contribute to pathological remodeling7,8.
Previous studies have demonstrated that spiny mouse extracellular matrix has different composition, structure, and properties that support regeneration in comparison to Mus musculus9,10. In muscle and heart injuries, FAPs have been noted to contribute to superior healing in spiny mouse11,12,13. Understanding the behavior and regulatory control of spiny mouse FAPs and stroma may shed light on the mechanism behind their regenerative capabilities. While FAPs are extensively studied in other animal models, such as in mus musculus14,15, currently, there are no published protocols for isolating spiny mouse FAPs. Developing such a protocol would fill a significant gap in the field and enable researchers to examine the cellular and molecular mechanisms underlying the spiny mouse&#39;s regenerative potential.
This protocol describes a robust and reproducible protocol for isolating, expanding, and differentiating skeletal and cardiac muscle FAPs from spiny mouse. The protocol described here yields high-quality single-cell suspensions that are suitable for fluorescence-activated cell sorting (FACS). By using rh-TGF&#946;1 or a compatible commercially available media, two methods widely used to differentiate mus musculus FAPs16, the sorted spiny FAPs maintain their capacity to differentiate along the fibrogenic lineage and adipogenic lineage, respectively (Figure 1).

<meta charset="utf8"></head><body>Pleins feux sur l’auteur : Rationaliser l’analyse cellulaire grâce à un protocole innovant pour l’étude de modèles de souris épineuses

Isolement, expansion et différenciation des progéniteurs fibro-adipogènes à partir du modèle de souris épineuses

Collecte de muscles squelettiques et cardiaques chez la souris épineuse pour isoler les progéniteurs fibro-adipogènes (PAF)

collection-skeletal-cardiac-muscles-from-spiny-mouse-for-isolating

Research

JoVE Journal

Developmental Biology

Collection of Skeletal and Cardiac Muscles from Spiny Mouse for Isolating Fibro-Adipogenic Progenitors (FAPs)

1.6K vues. Pour commencer, placez une souris euthanasiée sur une scène de dissection en position couchée, vaporisez soigneusement la fourrure de la souris avec de l’éthanol à 70%. Faites une incision verticale de deux à trois centimètres de long au niveau de la ligne médiane ventrale. Exposez la cage thoracique et le muscle abdominal.Ensuite, utilisez des ciseaux à tissus pour couper le muscle au niveau du processus xiphoïde et exposer le diaphragme. ...

Vidéo: Collecte de muscles squelettiques et cardiaques chez la souris épineuse pour isoler les progéniteurs fibro-adipogènes (PAF)

Fibro-Adipogenic Progenitor Isolation, Expansion, and Differentiation from the Spiny Mouse Model

Due to its exceptional repair program, the spiny mouse is an emerging research model for regenerative medicine. Fibro-adipogenic progenitors are tissue-resident cells that are able to differentiate into adipocytes, fibroblasts, and chondrocytes. Fibro-adipogenic progenitors are fundamental for orchestrating tissue regeneration as they are responsible for extracellular matrix remodeling after injury. This study focuses on investigating the specific role of fibro-adipogenic progenitors in spiny mouse cardiac repair and skeletal muscle regeneration. To this end, a protocol has been optimized for the purification of spiny mouse fibro-adipogenic progenitors by flow cytometry from enzymatically dissociated skeletal and cardiac muscle. The population obtained from this protocol is capable of expanding in vitro, and can be differentiated to myofibroblasts and adipocytes. This protocol offers a valuable tool for researchers to examine the distinctive properties of spiny mouse, and to compare them to the Mus musculus. This will provide insights that could advance the understanding of regenerative mechanisms in this intriguing model.

This protocol outlines the isolation of skeletal and cardiac muscle fibro-adipogenic progenitors (FAPs) from spiny mouse (Acomys) via enzymatic dissociation and fluorescence-activated cell sorting. The FAPs obtained from this protocol can be effectively expanded and differentiated to myofibroblasts and adipocytes.

Due to its exceptional repair program, the spiny mouse is an emerging research model for regenerative medicine. Fibro-adipogenic progenitors are ...

Recherche

Biologie du développement

Digestion of Mouse Skeletal and Cardiac Muscles for Fluorescence-Activated Cell Sorting and Culture of Fibro-Adipogenic Progenitors (FAPs)

Digestion des muscles squelettiques et cardiaques de souris pour le tri cellulaire activé par fluorescence et la culture de progéniteurs fibro-adipogènes (FAP)