Innovative methods for isolating highly purified mitochondria essential for biomedical studies
Mitochondria, being multifunctional and highly complex organelles, possess unique structures and exhibit heterogeneity. In recent decades, the isolation and purification of functional mitochondria have been instrumental for mitochondrial research. As mitochondrial research, including omics, advances...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2024-01-01
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| Series: | Mitochondrial Communications |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590279224000105 |
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| author | Yan Huang Xiangwaner Jin Yi Zhang Yanan Li Jinming Liu Yanjun Li |
| author_facet | Yan Huang Xiangwaner Jin Yi Zhang Yanan Li Jinming Liu Yanjun Li |
| author_sort | Yan Huang |
| collection | DOAJ |
| description | Mitochondria, being multifunctional and highly complex organelles, possess unique structures and exhibit heterogeneity. In recent decades, the isolation and purification of functional mitochondria have been instrumental for mitochondrial research. As mitochondrial research, including omics, advances, there is a growing demand for the isolation of highly purified mitochondria or individual mitochondria. This paper provides a comprehensive overview of the evolution of mitochondrial purification methods and introduces two innovative and improved techniques for isolating mitochondria from mouse cerebral cortex and in vitro cultured cells. The first method utilizes self-prepared magnetic beads conjugated with anti-TOMM20 antibody for the immunoisolation of highly purified intact mitochondria. The second method utilizes flow cytometry to isolate single mitochondria based on fluorescent protein labeling, allowing for the isolation of mitochondria from a highly heterogeneous population. We provide detailed protocols that aim to benefit the rapidly growing mitochondria research community in assessing mitochondrial function, especially at the single-organelle level. |
| format | Article |
| id | doaj-art-b7f492248ef9413da70e12602a8d9f8e |
| institution | Kabale University |
| issn | 2590-2792 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Mitochondrial Communications |
| spelling | doaj-art-b7f492248ef9413da70e12602a8d9f8e2025-08-20T03:31:40ZengKeAi Communications Co., Ltd.Mitochondrial Communications2590-27922024-01-012909910.1016/j.mitoco.2024.09.002Innovative methods for isolating highly purified mitochondria essential for biomedical studiesYan Huang0Xiangwaner Jin1Yi Zhang2Yanan Li3Jinming Liu4Yanjun Li5The Frontier Center for Cell Response, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, ChinaThe Frontier Center for Cell Response, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, ChinaThe Frontier Center for Cell Response, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, ChinaThe Frontier Center for Cell Response, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, ChinaThe Frontier Center for Cell Response, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, ChinaCorresponding author.; The Frontier Center for Cell Response, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, ChinaMitochondria, being multifunctional and highly complex organelles, possess unique structures and exhibit heterogeneity. In recent decades, the isolation and purification of functional mitochondria have been instrumental for mitochondrial research. As mitochondrial research, including omics, advances, there is a growing demand for the isolation of highly purified mitochondria or individual mitochondria. This paper provides a comprehensive overview of the evolution of mitochondrial purification methods and introduces two innovative and improved techniques for isolating mitochondria from mouse cerebral cortex and in vitro cultured cells. The first method utilizes self-prepared magnetic beads conjugated with anti-TOMM20 antibody for the immunoisolation of highly purified intact mitochondria. The second method utilizes flow cytometry to isolate single mitochondria based on fluorescent protein labeling, allowing for the isolation of mitochondria from a highly heterogeneous population. We provide detailed protocols that aim to benefit the rapidly growing mitochondria research community in assessing mitochondrial function, especially at the single-organelle level.http://www.sciencedirect.com/science/article/pii/S2590279224000105Mitochondrial purificationMagnetic beadFlow cytometryMitochondrion-associated protein aggregates (MAPAs)TOMM20 antibody |
| spellingShingle | Yan Huang Xiangwaner Jin Yi Zhang Yanan Li Jinming Liu Yanjun Li Innovative methods for isolating highly purified mitochondria essential for biomedical studies Mitochondrial Communications Mitochondrial purification Magnetic bead Flow cytometry Mitochondrion-associated protein aggregates (MAPAs) TOMM20 antibody |
| title | Innovative methods for isolating highly purified mitochondria essential for biomedical studies |
| title_full | Innovative methods for isolating highly purified mitochondria essential for biomedical studies |
| title_fullStr | Innovative methods for isolating highly purified mitochondria essential for biomedical studies |
| title_full_unstemmed | Innovative methods for isolating highly purified mitochondria essential for biomedical studies |
| title_short | Innovative methods for isolating highly purified mitochondria essential for biomedical studies |
| title_sort | innovative methods for isolating highly purified mitochondria essential for biomedical studies |
| topic | Mitochondrial purification Magnetic bead Flow cytometry Mitochondrion-associated protein aggregates (MAPAs) TOMM20 antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2590279224000105 |
| work_keys_str_mv | AT yanhuang innovativemethodsforisolatinghighlypurifiedmitochondriaessentialforbiomedicalstudies AT xiangwanerjin innovativemethodsforisolatinghighlypurifiedmitochondriaessentialforbiomedicalstudies AT yizhang innovativemethodsforisolatinghighlypurifiedmitochondriaessentialforbiomedicalstudies AT yananli innovativemethodsforisolatinghighlypurifiedmitochondriaessentialforbiomedicalstudies AT jinmingliu innovativemethodsforisolatinghighlypurifiedmitochondriaessentialforbiomedicalstudies AT yanjunli innovativemethodsforisolatinghighlypurifiedmitochondriaessentialforbiomedicalstudies |