A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts
Summary: This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to p...
| Published in: | STAR Protocols |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-12-01
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721005724 |
| _version_ | 1852818759615512576 |
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| author | Gabriella E. Farrugia Micheal A. McLellan Kate L. Weeks Aya Matsumoto Charles D. Cohen Crisdion Krstevski Taylah L. Gaynor Adam C. Parslow Julie R. McMullen Alexander R. Pinto |
| author_facet | Gabriella E. Farrugia Micheal A. McLellan Kate L. Weeks Aya Matsumoto Charles D. Cohen Crisdion Krstevski Taylah L. Gaynor Adam C. Parslow Julie R. McMullen Alexander R. Pinto |
| author_sort | Gabriella E. Farrugia |
| collection | DOAJ |
| container_title | STAR Protocols |
| description | Summary: This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to perfuse the heart with enzymes to dissociate cells. Myocytes and non-myocytes extracted using this protocol are separated by centrifugation and/or fluorescence-activated cell sorting for use in downstream applications including single-cell omics or other bio-molecular analyses.For complete details on the use and execution of this protocol, please refer to McLellan et al. (2020). |
| format | Article |
| id | doaj-art-e554ac6acc834cce8edb2df50ff09a7e |
| institution | Directory of Open Access Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2021-12-01 |
| publisher | Elsevier |
| record_format | Article |
| spelling | doaj-art-e554ac6acc834cce8edb2df50ff09a7e2025-08-19T20:33:06ZengElsevierSTAR Protocols2666-16672021-12-012410086610.1016/j.xpro.2021.100866A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse heartsGabriella E. Farrugia0Micheal A. McLellan1Kate L. Weeks2Aya Matsumoto3Charles D. Cohen4Crisdion Krstevski5Taylah L. Gaynor6Adam C. Parslow7Julie R. McMullen8Alexander R. Pinto9Baker Heart and Diabetes Institute, Melbourne, VIC 3004, AustraliaThe Jackson Laboratory, Bar Harbor, ME, USA; Tufts University, Graduate School of Biomedical Sciences, Boston, MA, USABaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Department of Diabetes, Central Clinical School, Monash University, Clayton, VIC 3800, Australia; Baker Department of Cardiometabolic Health, The University of Melbourne, Parkville, VIC 3010, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Baker Department of Cardiometabolic Health, The University of Melbourne, Parkville, VIC 3010, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Department of Diabetes, Central Clinical School, Monash University, Clayton, VIC 3800, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia; Department of Physiology and Department of Medicine Alfred Hospital, Monash University, Melbourne, VIC, Australia; Department of Physiology, Anatomy and Microbiology, La Trobe University, Bundoora, VIC, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia; Department of Physiology, Anatomy and Microbiology, La Trobe University, Bundoora, VIC, Australia; Corresponding authorSummary: This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to perfuse the heart with enzymes to dissociate cells. Myocytes and non-myocytes extracted using this protocol are separated by centrifugation and/or fluorescence-activated cell sorting for use in downstream applications including single-cell omics or other bio-molecular analyses.For complete details on the use and execution of this protocol, please refer to McLellan et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166721005724Cell BiologyCell isolationCell separation/fractionation |
| spellingShingle | Gabriella E. Farrugia Micheal A. McLellan Kate L. Weeks Aya Matsumoto Charles D. Cohen Crisdion Krstevski Taylah L. Gaynor Adam C. Parslow Julie R. McMullen Alexander R. Pinto A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts Cell Biology Cell isolation Cell separation/fractionation |
| title | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
| title_full | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
| title_fullStr | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
| title_full_unstemmed | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
| title_short | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
| title_sort | protocol for rapid and parallel isolation of myocytes and non myocytes from multiple mouse hearts |
| topic | Cell Biology Cell isolation Cell separation/fractionation |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721005724 |
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