Ischemia/reperfusion injury of primary porcine cardiomyocytes in a low-shear microfluidic culture and analysis device.
Ischemia/reperfusion injury of primary porcine cardiomyocytes in a low-shear microfluidic culture and analysis device.
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DOI：
10.1039/c0an00845a
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发表时间：
2011-09-07
期刊：
影响因子：
--
通讯作者：
Pappas D
中科院分区：
文献类型：
--
作者：
Khanal G;Chung K;Solis-Wever X;Johnson B;Pappas D
在低剪切微流控培养芯片中诱导原代猪心肌细胞缺血/再灌注（I/R）损伤。该芯片能够维持心肌细胞培养，并通过使细胞经历持续3-4小时的缺氧期，然后是常氧来诱导I/R损伤。使用MitoTracker Red测定线粒体膜电位，以跟踪线粒体去极化，即细胞凋亡的最早阶段。细胞粘附和形态也同时测定与荧光测量。膜电位的变化比以前报道的要早，线粒体早在缺血2小时就去极化了。具有去极化线粒体的细胞被认为是凋亡的。在每个时间范围的38-61个细胞中，发现在缺血两小时时凋亡细胞的分数与对照样品（3%）相似，与对照样品中的0%相比，在缺血期结束时增加至22%。细胞的形态学分析表明，4小时的缺血再灌注在2小时内恢复芯片的氧气产生水泡细胞。这种方法是一种通用的心肌细胞应激的方法，在未来的工作中，额外的分析探针可以被纳入心肌细胞凋亡的多分析物测定。
Ischemia/reperfusion (I/R) injury was induced in primary porcine cardiomyocytes in a low-shear microfluidic culture chip. The chip was capable of sustaining cardiomyocyte culture and inducing I/R injury by subjecting the cells to periods of hypoxia lasting 3-4 hours followed by normoxia. Mitochondrial membrane potential was assayed using MitoTracker Red to follow mitochondrial depolarization, the earliest stage of apoptosis. Cell adhesion and morphology were also determined simultaneously with fluorescence measurements. Changes in membrane potential were observed earlier than previously reported, with mitochondria becoming depolarized as early as 2 hours into the ischemia period. The cells with depolarized mitochondria were deemed apoptotic. Out of 38-61 cells per time frame, the fraction of apoptotic cells was found to be similar to control samples (3%) at two hours of ischemia, which increased up to 22% at the end of ischemia period as compared to 0% in the control samples. Morphological analysis of cells showed that 4 hours of ischemia followed by reperfusion produced blebbing cells within 2 hours of restoring oxygen to the chip. This approach is a versatile method for cardiomyocyte stress, and in future work additional analytical probes can be incorporated for a multi-analyte assays of cardiomyocyte apoptosis.
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影响因子：
3.9
作者：
Liu, Ke;Pitchimani, Rajasekar;Pappas, Dimitri
通讯作者：
Pappas, Dimitri
影响因子：
5
作者：
Holly, TA;Drincic, A;Cryns, VL
通讯作者：
Cryns, VL
微芯片电泳-激光诱导荧光同时测定凋亡细胞线粒体区室中谷胱甘肽和过氧化氢的有效方法
影响因子：
7.4
作者：
Chen, Zhenzhen;Li, Qingling;Tang, Bo
通讯作者：
Tang, Bo
DOI：
10.1152/ajpheart.00777.2002
发表时间：
2003-02-01
影响因子：
4.8
作者：
Lee, P;Sata, M;Kitsis, RN
通讯作者：
Kitsis, RN
影响因子：
6.1
作者：
Oppegard, Shawn C.;Blake, Alexander J.;Eddington, David T.
通讯作者：
Eddington, David T.