应用 ECIS 评估 FCCP 诱导的 MSC 微运动变化及其对创伤愈合迁移的影响。
Application of ECIS to Assess FCCP-Induced Changes of MSC Micromotion and Wound Healing Migration.
机构信息
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.
Division of Endocrinology and Metabolism, Department of Internal Medicine, Tri-Service General Hospital Songshan Branch, National Defense Medical Center, Taipei 11490, Taiwan.
出版信息
Sensors (Basel). 2019 Jul 21;19(14):3210. doi: 10.3390/s19143210.
Electric cell-substrate impedance sensing (ECIS) is an emerging technique for sensitively monitoring morphological changes of adherent cells in tissue culture. In this study, human mesenchymal stem cells (hMSCs) were exposed to different concentrations of carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) for 20 h and their subsequent concentration-dependent responses in micromotion and wound healing migration were measured by ECIS. FCCP disrupts ATP synthesis and results in a decrease in cell migration rates. To detect the change of cell micromotion in response to FCCP challenge, time-series resistances of cell-covered electrodes were monitored and the values of variance were calculated to verify the difference. While Seahorse XF-24 extracellular flux analyzer can detect the effect of FCCP at 3 μM concentration, the variance calculation of the time-series resistances measured at 4 kHz can detect the effect of FCCP at concentrations as low as 1 μM. For wound healing migration, the recovery resistance curves were fitted by sigmoid curve and the hill slope showed a concentration-dependent decline from 0.3 μM to 3 μM, indicating a decrease in cell migration rate. Moreover, dose dependent incline of the inflection points from 0.3 μM to 3 μM FCCP implied the increase of the half time for wound recovery migration. Together, our results demonstrate that partial uncoupling of mitochondrial oxidative phosphorylation reduces micromotion and wound healing migration of hMSCs. The ECIS method used in this study offers a simple and sensitive approach to investigate stem cell migration and its regulation by mitochondrial dynamics.
电细胞-基底阻抗感应（ECIS）是一种新兴的技术，可灵敏地监测组织培养中贴壁细胞的形态变化。在这项研究中，将人骨髓间充质干细胞（hMSC）暴露于不同浓度的羰基氰化物 4-（三氟甲氧基）苯腙（FCCP）20 小时，并通过 ECIS 测量其随后的浓度依赖性微运动和伤口愈合迁移反应。FCCP 破坏 ATP 合成，导致细胞迁移率降低。为了检测细胞对 FCCP 刺激的微运动变化，监测细胞覆盖电极的时间序列电阻，并计算方差值以验证差异。虽然 Seahorse XF-24 细胞外通量分析仪可以检测 3 μM 浓度的 FCCP 作用，但在 4 kHz 测量的时间序列电阻的方差计算可以检测低至 1 μM 浓度的 FCCP 作用。对于伤口愈合迁移，通过 S 型曲线拟合恢复电阻曲线，斜率显示出从 0.3 μM 到 3 μM 的浓度依赖性下降，表明细胞迁移率降低。此外，从 0.3 μM 到 3 μM FCCP 的拐点的剂量依赖性倾斜表明伤口恢复迁移的半衰期增加。总之，我们的结果表明，线粒体氧化磷酸化的部分解偶联降低了 hMSC 的微运动和伤口愈合迁移。本研究中使用的 ECIS 方法提供了一种简单灵敏的方法来研究干细胞迁移及其受线粒体动力学的调节。
Zotero 插件