Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality

Abstract

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

分享

查看原文 本刊更多论文

分离胰腺腺泡的非偶联呼吸稳定性作为细胞活力的新功能测试

背景。细胞活力的评估在细胞研究中是至关重要的。检测细胞膜完整性是评估细胞活力的传统方法。线粒体功能能力与质膜完整性和整体细胞健康密切相关。本研究旨在探讨分离胰腺腺泡细胞线粒体适应能力的任何方面是否与分离胰腺腺泡细胞制剂的质量有关，通过染料排除法确定。材料与方法。实验以体重250 ~ 300 g的雄性Wistar大鼠为实验对象。用胶原酶获得分离胰腺腺泡悬液。用Clark氧电极测定大鼠离体胰腺腺泡的耗氧量。记录离体胰腺腺泡细胞的基础呼吸约2分钟，然后用浓度为0.5 ~ 2 μM的FCCP检测线粒体适应能力。非耦合呼吸稳定性计算为高、低FCCP浓度下呼吸速率的比值。台盼蓝染色评估质膜完整性。本研究共使用了74种分离胰腺腺泡制剂。结果。在所有实验中，92 - 99%的胰腺腺泡细胞呈阴性台盼蓝染色，表明质膜完整。基础呼吸速率和最大不耦合呼吸速率不受台盼阴性细胞比例的影响。然而，当暴露于高浓度FCCP(2µM)时，小于95%质膜完整性的腺泡菌制剂的解耦呼吸速率显著降低，表明解耦呼吸的稳定性降低。结论。研究结果表明，非偶联呼吸的稳定性可以作为一种新的代谢功能测试来补充现有的评估细胞活力的方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。