Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality
Background. Assessment of cell viability is crucial in cell studies. Testing plasma membrane integrity is a traditional approach of evaluating cell viability. Mitochondrial functional capacity closely correlates with plasma membrane integrity and overall cell health. This study aimed to investigate...
| Principais autores: | , , , , |
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| Formato: | Artigo |
| Idioma: | English |
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Львівський національний університет імені Івана Франка
2023-09-01
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| coleção: | Біологічні студії |
| Assuntos: | |
| Acesso em linha: | http://publications.lnu.edu.ua/journals/index.php/biology/article/view/3356 |
| _version_ | 1827806220002525184 |
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| author | Anastasiia Zub Bohdan V. Manko Bohdan O. Manko Volodymyr Manko Andriy Babsky |
| author_facet | Anastasiia Zub Bohdan V. Manko Bohdan O. Manko Volodymyr Manko Andriy Babsky |
| author_sort | Anastasiia Zub |
| collection | DOAJ |
| description | Background. Assessment of cell viability is crucial in cell studies. Testing plasma membrane integrity is a traditional approach of evaluating cell viability. Mitochondrial functional capacity closely correlates with plasma membrane integrity and overall cell health. This study aimed to investigate whether any aspect of mitochondrial adaptive capacity in isolated pancreatic acini is associated with the quality of isolated pancreatic acini preparations, as determined by the dye exclusion method.
Materials and Methods. Experiments were carried out on male Wistar rats weighing 250–300 g. A suspension of isolated pancreatic acini was obtained using collagenase. The rate of oxygen consumption of rat isolated pancreatic acini was measured with Clark oxygen electrode. Basal respiration of isolated pancreatic acini was recorded for approximately 2 min. Afterwards, the mitochondrial adaptive capacity was examined using FCCP in concentrations from 0.5 to 2 μM. Uncoupled respiratory stability was calculated as a ratio of respiration rate at high and low FCCP concentrations. Plasma membrane integrity was assessed with trypan blue staining. A total of 74 preparations of isolated pancreatic acini were used in this study.
Results. In all experiments, 92–99 % of pancreatic acinar cells exhibited negative trypan blue staining, indicating intact plasma membranes. The basal and maximal uncoupled respiration rates were not affected by the fraction of trypan-negative cells. However, acini preparations with <less than 95 % plasma membrane integrity had significantly lower uncoupled respiration rates when exposed to a high concentration of FCCP (2 µM), indicating reduced stability of uncoupled respiration.
Conclusions. Results of the study suggest that the stability of uncoupled respiration can serve as a novel metabolic functional test to complement the existing methods for assessing cell vitality. |
| first_indexed | 2024-03-11T21:38:04Z |
| format | Article |
| id | doaj.art-b2876792e3974fb6b2479b648a9837bf |
| institution | Directory Open Access Journal |
| issn | 1996-4536 2311-0783 |
| language | English |
| last_indexed | 2024-03-11T21:38:04Z |
| publishDate | 2023-09-01 |
| publisher | Львівський національний університет імені Івана Франка |
| record_format | Article |
| series | Біологічні студії |
| spelling | doaj.art-b2876792e3974fb6b2479b648a9837bf2023-09-26T17:04:42ZengЛьвівський національний університет імені Івана ФранкаБіологічні студії1996-45362311-07832023-09-0117324325210.30970/sbi.1703.735Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitalityAnastasiia Zub0https://orcid.org/0000-0002-9335-0928Bohdan V. Manko1https://orcid.org/0000-0001-9762-4263Bohdan O. Manko2https://orcid.org/0000-0002-6905-8580Volodymyr Manko3https://orcid.org/0000-0002-4406-1732Andriy Babsky4https://orcid.org/0000-0001-5559-0456Ivan Franko National University of LvivIvan Franko National University of LvivIvan Franko National University of LvivIvan Franko National University of LvivIvan Franko National University of LvivBackground. Assessment of cell viability is crucial in cell studies. Testing plasma membrane integrity is a traditional approach of evaluating cell viability. Mitochondrial functional capacity closely correlates with plasma membrane integrity and overall cell health. This study aimed to investigate whether any aspect of mitochondrial adaptive capacity in isolated pancreatic acini is associated with the quality of isolated pancreatic acini preparations, as determined by the dye exclusion method. Materials and Methods. Experiments were carried out on male Wistar rats weighing 250–300 g. A suspension of isolated pancreatic acini was obtained using collagenase. The rate of oxygen consumption of rat isolated pancreatic acini was measured with Clark oxygen electrode. Basal respiration of isolated pancreatic acini was recorded for approximately 2 min. Afterwards, the mitochondrial adaptive capacity was examined using FCCP in concentrations from 0.5 to 2 μM. Uncoupled respiratory stability was calculated as a ratio of respiration rate at high and low FCCP concentrations. Plasma membrane integrity was assessed with trypan blue staining. A total of 74 preparations of isolated pancreatic acini were used in this study. Results. In all experiments, 92–99 % of pancreatic acinar cells exhibited negative trypan blue staining, indicating intact plasma membranes. The basal and maximal uncoupled respiration rates were not affected by the fraction of trypan-negative cells. However, acini preparations with <less than 95 % plasma membrane integrity had significantly lower uncoupled respiration rates when exposed to a high concentration of FCCP (2 µM), indicating reduced stability of uncoupled respiration. Conclusions. Results of the study suggest that the stability of uncoupled respiration can serve as a novel metabolic functional test to complement the existing methods for assessing cell vitality.http://publications.lnu.edu.ua/journals/index.php/biology/article/view/3356pancreasacinar cellsviabilityuncoupled respiration |
| spellingShingle | Anastasiia Zub Bohdan V. Manko Bohdan O. Manko Volodymyr Manko Andriy Babsky Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality Біологічні студії pancreas acinar cells viability uncoupled respiration |
| title | Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality |
| title_full | Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality |
| title_fullStr | Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality |
| title_full_unstemmed | Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality |
| title_short | Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality |
| title_sort | uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality |
| topic | pancreas acinar cells viability uncoupled respiration |
| url | http://publications.lnu.edu.ua/journals/index.php/biology/article/view/3356 |
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