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Updated: Aug 6, 2025
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Preparation of 3D Decellularized Matrices from Fetal Mouse Skeletal Muscle for Cell Culture
Published on: March 3, 2023
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Preparation of 3D Decellularized Matrices from Fetal Mouse Skeletal Muscle for Cell Culture
Pedro Gameiro Dos Santos1, Ana Rita Soares2, Sólveig Thorsteinsdóttir2
1Centre for Ecology, Evolution and Environmental Changes and Global Change & Sustainability Institute, Department of Animal Biology, Faculty of Sciences, University of Lisbon; pgsantos@fc.ul.pt.
Journal of Visualized Experiments : Jove
|March 20, 2023
Summary
Researchers developed a 3D model using decellularized extracellular matrix (dECM) from fetal mouse muscle. This model supports muscle cell growth and differentiation, aiding the study of congenital muscular dystrophy and other ECM-related diseases.
Area of Science:
- Biomaterials Science
- Cell Biology
- Developmental Biology
Background:
- The extracellular matrix (ECM) is vital for cell structure and signaling, but 2D cultures inadequately mimic its in vivo complexity.
- Dysfunctional ECM and cell-ECM interactions contribute to diseases like LAMA2-congenital muscular dystrophy (LAMA2-CMD).
- LAMA2-CMD, caused by laminin deficiency, leads to severe hypotonia during fetal development.
Purpose of the Study:
- To create a 3D in vitro model of fetal muscle ECM for studying cell-ECM interactions.
- To mimic the native microenvironment for investigating muscle development and disease onset.
- To provide a platform for understanding LAMA2-CMD and other skeletal muscle diseases.
Main Methods:
- Dissection of deep back muscles from E18.5 mouse fetuses.
- Decellularization of muscle tissue using hypotonic buffer, anionic detergent, and DNase.
- Seeding C2C12 myoblasts onto the resulting decellularized matrices (dECMs).
Main Results:
- Decellularized matrices (dECMs) retained key ECM proteins including laminin α2, fibronectin, collagen I, and collagen IV.
- C2C12 myoblasts successfully colonized, proliferated, and differentiated within the dECMs.
- Seeded cells actively produced ECM proteins, remodeling their microenvironment.
Conclusions:
- The developed 3D dECM model effectively supports muscle cell behavior, mimicking the native environment.
- This platform offers a promising approach to study LAMA2-CMD pathogenesis during fetal myogenesis.
- The model can be adapted for investigating other skeletal muscle diseases linked to ECM communication deficits.
