Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies
The aim of this work was to develop a cryopreservation method of small liver biopsies for in situ mitochondrial function assessment. Herein we describe a detailed protocol for tissue collection, cryopreservation, high-resolution respirometry using complex I and II substrates, calculation and interpr...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-07-01
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| Series: | Redox Biology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231718301472 |
| _version_ | 1811226243832479744 |
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| author | Mercedes García-Roche Alberto Casal Mariana Carriquiry Rafael Radi Celia Quijano Adriana Cassina |
| author_facet | Mercedes García-Roche Alberto Casal Mariana Carriquiry Rafael Radi Celia Quijano Adriana Cassina |
| author_sort | Mercedes García-Roche |
| collection | DOAJ |
| description | The aim of this work was to develop a cryopreservation method of small liver biopsies for in situ mitochondrial function assessment. Herein we describe a detailed protocol for tissue collection, cryopreservation, high-resolution respirometry using complex I and II substrates, calculation and interpretation of respiratory parameters. Liver biopsies from cow and rat were sequentially frozen in a medium containing dimethylsulfoxide as cryoprotectant and stored for up to 3 months at −80 °C. Oxygen consumption rate studies of fresh and cryopreserved samples revealed that most respiratory parameters remained unchanged. Additionally, outer mitochondrial membrane integrity was assessed adding cytochrome c, proving that our cryopreservation method does not harm mitochondrial structure. In sum, we present a reliable way to cryopreserve small liver biopsies without affecting mitochondrial function. Our protocol will enable the transport and storage of samples, extending and facilitating mitochondrial function analysis of liver biopsies. Keywords: Cryopreservation, Mitochondria, Biopsy, Oxygen consumption rate, High-resolution respirometry, Mitochondrial function |
| first_indexed | 2024-04-12T09:21:45Z |
| format | Article |
| id | doaj.art-9713fd500363411bad5f521957837393 |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-04-12T09:21:45Z |
| publishDate | 2018-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-9713fd500363411bad5f5219578373932022-12-22T03:38:37ZengElsevierRedox Biology2213-23172018-07-0117207212Respiratory analysis of coupled mitochondria in cryopreserved liver biopsiesMercedes García-Roche0Alberto Casal1Mariana Carriquiry2Rafael Radi3Celia Quijano4Adriana Cassina5Center for Free Radical and Biomedical Research (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Uruguay; Departamento de Producción Animal y Pasturas, Facultad de Agronomía, Universidad de la República, UruguayDepartamento de Producción Animal y Pasturas, Facultad de Agronomía, Universidad de la República, UruguayDepartamento de Producción Animal y Pasturas, Facultad de Agronomía, Universidad de la República, UruguayCenter for Free Radical and Biomedical Research (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, UruguayCenter for Free Radical and Biomedical Research (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Uruguay; Correspondence to: Center for Free Radical and Biomedical Research (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Avenida General Flores 2125, Montevideo 11800, Uruguay.Center for Free Radical and Biomedical Research (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Uruguay; Correspondence to: Center for Free Radical and Biomedical Research (CEINBIO) and Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Avenida General Flores 2125, Montevideo 11800, Uruguay.The aim of this work was to develop a cryopreservation method of small liver biopsies for in situ mitochondrial function assessment. Herein we describe a detailed protocol for tissue collection, cryopreservation, high-resolution respirometry using complex I and II substrates, calculation and interpretation of respiratory parameters. Liver biopsies from cow and rat were sequentially frozen in a medium containing dimethylsulfoxide as cryoprotectant and stored for up to 3 months at −80 °C. Oxygen consumption rate studies of fresh and cryopreserved samples revealed that most respiratory parameters remained unchanged. Additionally, outer mitochondrial membrane integrity was assessed adding cytochrome c, proving that our cryopreservation method does not harm mitochondrial structure. In sum, we present a reliable way to cryopreserve small liver biopsies without affecting mitochondrial function. Our protocol will enable the transport and storage of samples, extending and facilitating mitochondrial function analysis of liver biopsies. Keywords: Cryopreservation, Mitochondria, Biopsy, Oxygen consumption rate, High-resolution respirometry, Mitochondrial functionhttp://www.sciencedirect.com/science/article/pii/S2213231718301472 |
| spellingShingle | Mercedes García-Roche Alberto Casal Mariana Carriquiry Rafael Radi Celia Quijano Adriana Cassina Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies Redox Biology |
| title | Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies |
| title_full | Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies |
| title_fullStr | Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies |
| title_full_unstemmed | Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies |
| title_short | Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies |
| title_sort | respiratory analysis of coupled mitochondria in cryopreserved liver biopsies |
| url | http://www.sciencedirect.com/science/article/pii/S2213231718301472 |
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