Animal Physiology and Reproduction Division, ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125001, India.
Animal Physiology and Reproduction Division, ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana 125001, India.
Tissue Cell. 2023 Jun;82:102067. doi: 10.1016/j.tice.2023.102067. Epub 2023 Mar 11.
Cryopreservation commonly decreases the cellular functionality and post-thaw viability of cells. Reactive oxygen species (ROS) generated during cryopreservation degrade mitochondrial activity and promote the release of cytochrome C which activates caspases required for apoptosis. Antioxidants have the potential to improve the recovery efficiency of cells by reducing ROS production and maintaining mitochondrial membrane potential (MMP). The present study was conducted to explore the role of MitoQ, a derivative of coenzyme Q10 on cryopreserved fibroblasts derived from buffalo skin. To achieve our goal, buffalo skin fibroblasts were treated with varying concentrations of MitoQ (0, 0.1, 0.5, 1, 2, and 10 μM) for 24, 48, and 72 h. The MMP, ROS generation, cell viability was measured by flow cytometry. Furthermore, expression of genes related to mitochondrial oxidative stress (NRF2, GPX, and SOD), apoptosis (BAK and caspase 3) and cell proliferation (AKT) were also assessed. The results showed that over a period of 72 h lower concentrations of MitoQ (0.1-0.5 μM) decrease the ROS production, improves MMP and cell viability whilst the high concentration of MitoQ (2-10 μM) increased the oxidative damage to the cells. Taken together, our study provide important insights into the novel role of MitoQ in cryopreserved buffalo skin fibroblasts. In conclusion, we demonstrated the dose-dependent functional role of MitoQ on cryopreserved fibroblasts for improving post-thaw cell viability and cellular function.
冷冻保存通常会降低细胞的功能和冻后活力。冷冻保存过程中产生的活性氧 (ROS) 会降低线粒体活性，并促进细胞色素 C 的释放，从而激活细胞凋亡所需的半胱天冬酶。抗氧化剂通过减少 ROS 生成和维持线粒体膜电位 (MMP) ，有可能提高细胞的回收效率。本研究旨在探索 MitoQ（辅酶 Q10 的衍生物）在水牛皮肤来源的冷冻保存成纤维细胞中的作用。为了实现我们的目标，用不同浓度的 MitoQ（0、0.1、0.5、1、2 和 10 μM）处理水牛皮肤成纤维细胞 24、48 和 72 h。通过流式细胞术测量 MMP、ROS 生成和细胞活力。此外，还评估了与线粒体氧化应激（NRF2、GPX 和 SOD）、凋亡（BAK 和 caspase 3）和细胞增殖（AKT）相关的基因的表达。结果表明，在 72 h 的时间内，较低浓度的 MitoQ（0.1-0.5 μM）可减少 ROS 生成，提高 MMP 和细胞活力，而较高浓度的 MitoQ（2-10 μM）会增加细胞的氧化损伤。总之，我们的研究为 MitoQ 在冷冻保存的水牛皮肤成纤维细胞中的新作用提供了重要的见解。综上所述，我们证明了 MitoQ 在提高冻后细胞活力和细胞功能方面对冷冻保存成纤维细胞具有剂量依赖性的功能作用。
