Reporter-Based Isolation of Developmental Myogenic Progenitors
The formation and activity of mammalian tissues entail finely regulated processes, involving the concerted organization and interaction of multiple cell types. In recent years the prospective isolation of distinct progenitor and stem cell populations has become a powerful tool in the hands of develo...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-04-01
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| Series: | Frontiers in Physiology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/article/10.3389/fphys.2018.00352/full |
| _version_ | 1831729077763964928 |
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| author | Eyemen Kheir Eyemen Kheir Gabriella Cusella Gabriella Cusella Graziella Messina Giulio Cossu Stefano Biressi Stefano Biressi |
| author_facet | Eyemen Kheir Eyemen Kheir Gabriella Cusella Gabriella Cusella Graziella Messina Giulio Cossu Stefano Biressi Stefano Biressi |
| author_sort | Eyemen Kheir |
| collection | DOAJ |
| description | The formation and activity of mammalian tissues entail finely regulated processes, involving the concerted organization and interaction of multiple cell types. In recent years the prospective isolation of distinct progenitor and stem cell populations has become a powerful tool in the hands of developmental biologists and has rendered the investigation of their intrinsic properties possible. In this protocol, we describe how to purify progenitors with different lineage history and degree of differentiation from embryonic and fetal skeletal muscle by fluorescence-activated cell sorting (FACS). The approach takes advantage of a panel of murine strains expressing fluorescent reporter genes specifically in the myogenic progenitors. We provide a detailed description of the dissection procedures and of the enzymatic dissociation required to maximize the yield of mononucleated cells for subsequent FACS-based purification. The procedure takes ~6–7 h to complete and allows for the isolation and the subsequent molecular and phenotypic characterization of developmental myogenic progenitors. |
| first_indexed | 2024-12-21T07:01:47Z |
| format | Article |
| id | doaj.art-1917eb44b0014f57a64b1a320460f98b |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-21T07:01:47Z |
| publishDate | 2018-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-1917eb44b0014f57a64b1a320460f98b2022-12-21T19:12:11ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2018-04-01910.3389/fphys.2018.00352350792Reporter-Based Isolation of Developmental Myogenic ProgenitorsEyemen Kheir0Eyemen Kheir1Gabriella Cusella2Gabriella Cusella3Graziella Messina4Giulio Cossu5Stefano Biressi6Stefano Biressi7Centre for Integrative Biology (CIBIO), University of Trento, Trento, ItalyDulbecco Telethon Institute, University of Trento, Trento, ItalyHuman Anatomy Unit, Department of Public Health, Experimental Medicine and Forensic, University of Pavia, Pavia, ItalyCenter for Health Technologies, University of Pavia, Pavia, ItalyDepartment of Biosciences, University of Milan, Milan, ItalyDivision of Cell Matrix Biology and Regenerative Medicine, Manchester Academic Health Science Centre, University of Manchester, Manchester, United KingdomCentre for Integrative Biology (CIBIO), University of Trento, Trento, ItalyDulbecco Telethon Institute, University of Trento, Trento, ItalyThe formation and activity of mammalian tissues entail finely regulated processes, involving the concerted organization and interaction of multiple cell types. In recent years the prospective isolation of distinct progenitor and stem cell populations has become a powerful tool in the hands of developmental biologists and has rendered the investigation of their intrinsic properties possible. In this protocol, we describe how to purify progenitors with different lineage history and degree of differentiation from embryonic and fetal skeletal muscle by fluorescence-activated cell sorting (FACS). The approach takes advantage of a panel of murine strains expressing fluorescent reporter genes specifically in the myogenic progenitors. We provide a detailed description of the dissection procedures and of the enzymatic dissociation required to maximize the yield of mononucleated cells for subsequent FACS-based purification. The procedure takes ~6–7 h to complete and allows for the isolation and the subsequent molecular and phenotypic characterization of developmental myogenic progenitors.http://journal.frontiersin.org/article/10.3389/fphys.2018.00352/fullembryonic myoblastsfetal myoblatsFACSreporter linesmyf5 |
| spellingShingle | Eyemen Kheir Eyemen Kheir Gabriella Cusella Gabriella Cusella Graziella Messina Giulio Cossu Stefano Biressi Stefano Biressi Reporter-Based Isolation of Developmental Myogenic Progenitors Frontiers in Physiology embryonic myoblasts fetal myoblats FACS reporter lines myf5 |
| title | Reporter-Based Isolation of Developmental Myogenic Progenitors |
| title_full | Reporter-Based Isolation of Developmental Myogenic Progenitors |
| title_fullStr | Reporter-Based Isolation of Developmental Myogenic Progenitors |
| title_full_unstemmed | Reporter-Based Isolation of Developmental Myogenic Progenitors |
| title_short | Reporter-Based Isolation of Developmental Myogenic Progenitors |
| title_sort | reporter based isolation of developmental myogenic progenitors |
| topic | embryonic myoblasts fetal myoblats FACS reporter lines myf5 |
| url | http://journal.frontiersin.org/article/10.3389/fphys.2018.00352/full |
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