Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system
Decellularisation of skeletal muscle provides a system to study the interactions of myoblasts with muscle extracellular matrix (ECM). This study describes the efficient decellularisation of quadriceps muscle with the retention of matrix components and the use of this matrix for myoblast proliferatio...
| Main Authors: | , , , , , |
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| Format: | Journal Article |
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Public Library of Science
2015
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| Online Access: | http://hdl.handle.net/20.500.11937/6742 |
| _version_ | 1848745164308217856 |
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| author | Chaturvedi, Vishal Dye, Danielle Kinnear, Bev Van Kuppevelt, T. Grounds, M. Coombe, Deidre |
| author_facet | Chaturvedi, Vishal Dye, Danielle Kinnear, Bev Van Kuppevelt, T. Grounds, M. Coombe, Deidre |
| author_sort | Chaturvedi, Vishal |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | Decellularisation of skeletal muscle provides a system to study the interactions of myoblasts with muscle extracellular matrix (ECM). This study describes the efficient decellularisation of quadriceps muscle with the retention of matrix components and the use of this matrix for myoblast proliferation and differentiation under serum free culture conditions. Three decellularisation approaches were examined; the most effective was phospholipase A2 treatment, which removed cellular material while maximizing the retention of ECM components. Decellularised muscle matrices were then solubilized and used as substrates for C2C12 mouse myoblast serum free cultures. The muscle matrix supported myoblast proliferation and differentiation equally as well as collagen and fibronectin. Immunofluorescence analyses revealed that myoblasts seeded on muscle matrix and fibronectin differentiated to form long, well-aligned myotubes, while myoblasts seeded on collagen were less organized. qPCR analyses showed a time dependent increase in genes involved in skeletal muscle differentiation and suggested that muscle-derived matrix may stimulate an increased rate of differentiation compared to collagen and fibronectin. Decellularized whole muscle three-dimensional scaffolds also supported cell adhesion and spreading, with myoblasts aligning along specific tracts of matrix proteins within the scaffolds. Thus, under serum free conditions, intact acellular muscle matrices provided cues to direct myoblast adhesion and migration. In addition, myoblasts were shown to rapidly secrete and organise their own matrix glycoproteins to create a localized ECM microenvironment. This serum free culture system has revealed that the correct muscle ECM facilitates more rapid cell organisation and differentiation than single matrix glycoprotein substrates. |
| first_indexed | 2025-11-14T06:13:00Z |
| format | Journal Article |
| id | curtin-20.500.11937-6742 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T06:13:00Z |
| publishDate | 2015 |
| publisher | Public Library of Science |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-67422017-09-13T14:34:28Z Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system Chaturvedi, Vishal Dye, Danielle Kinnear, Bev Van Kuppevelt, T. Grounds, M. Coombe, Deidre Decellularisation of skeletal muscle provides a system to study the interactions of myoblasts with muscle extracellular matrix (ECM). This study describes the efficient decellularisation of quadriceps muscle with the retention of matrix components and the use of this matrix for myoblast proliferation and differentiation under serum free culture conditions. Three decellularisation approaches were examined; the most effective was phospholipase A2 treatment, which removed cellular material while maximizing the retention of ECM components. Decellularised muscle matrices were then solubilized and used as substrates for C2C12 mouse myoblast serum free cultures. The muscle matrix supported myoblast proliferation and differentiation equally as well as collagen and fibronectin. Immunofluorescence analyses revealed that myoblasts seeded on muscle matrix and fibronectin differentiated to form long, well-aligned myotubes, while myoblasts seeded on collagen were less organized. qPCR analyses showed a time dependent increase in genes involved in skeletal muscle differentiation and suggested that muscle-derived matrix may stimulate an increased rate of differentiation compared to collagen and fibronectin. Decellularized whole muscle three-dimensional scaffolds also supported cell adhesion and spreading, with myoblasts aligning along specific tracts of matrix proteins within the scaffolds. Thus, under serum free conditions, intact acellular muscle matrices provided cues to direct myoblast adhesion and migration. In addition, myoblasts were shown to rapidly secrete and organise their own matrix glycoproteins to create a localized ECM microenvironment. This serum free culture system has revealed that the correct muscle ECM facilitates more rapid cell organisation and differentiation than single matrix glycoprotein substrates. 2015 Journal Article http://hdl.handle.net/20.500.11937/6742 10.1371/journal.pone.0127675 Public Library of Science fulltext |
| spellingShingle | Chaturvedi, Vishal Dye, Danielle Kinnear, Bev Van Kuppevelt, T. Grounds, M. Coombe, Deidre Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| title | Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| title_full | Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| title_fullStr | Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| title_full_unstemmed | Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| title_short | Interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| title_sort | interactions between skeletal muscle myoblasts and their extracellular matrix revealed by a serum free culture system |
| url | http://hdl.handle.net/20.500.11937/6742 |