Advancing practical usage of microtechnology: a study of the functional consequences of dielectrophoresis on neural stem cells.
Advancing practical usage of microtechnology: a study of the functional consequences of dielectrophoresis on neural stem cells.
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DOI：
10.1039/c2ib20171b
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发表时间：
2012-10
期刊：
影响因子：
--
通讯作者：
Flanagan LA
中科院分区：
文献类型：
--
作者：
Lu J;Barrios CA;Dickson AR;Nourse JL;Lee AP;Flanagan LA
微尺度工程、微流体和交流电动力学(如介电泳法)的结合产生了新的微系统，使细胞表型、功能和生理的定量分析成为可能。这些系统越来越多地被用于评估不同类型的细胞，如干细胞，因此彻底评估系统本身是否影响细胞功能变得至关重要。例如，工程微系统已被用于研究神经干细胞/祖细胞(NSPC)，由于其治疗中枢神经系统疾病和损伤的潜力而引起人们的兴趣。介电分析(DEP)微系统分析表明，具有不同命运潜力的未标记NSPC具有以前未被识别的区分电生理特性，这表明DEP微系统可以在不使用细胞类型特定标记的情况下分离NSPC。为了评估DEP分选对NSPC的潜在影响，我们调查了不同时间的电场暴露是否影响悬液中NSPC的存活、增殖或命运潜力。我们发现，短期的DEP暴露(1分钟或更短)对NSPC的存活、增殖和分化所揭示的命运潜能没有影响。此外，NSPC的增殖(通过DNA合成和细胞周期动力学测量)和命运潜力不受任何DEP暴露时间的影响(最长为30min)。然而，长时间(>；5分钟)暴露于接近交叉频率(50-100 kHz)的频率会导致NSPC存活率下降(30分钟后最大细胞损失率约为30%)。基于对悬浮细胞的实验观察和数学模拟，我们发现接近交叉频率的频率会产生诱导的跨膜电位，导致细胞肿胀和破裂。这与贴壁细胞的情况相反，因为负的DEP频率低于交叉频率会对这些细胞产生最高的诱导跨膜电位和损伤。我们阐明了DEP对贴壁细胞和悬浮细胞的对比效应，这与细胞在电场中的位置和距离电极特定距离的电场强度有关。电极构型的建模预测了最佳设计，以在限制诱导的跨膜电位的同时通过DEP诱导细胞移动。我们发现DEP电场在短时间内对悬浮液中的干细胞无害，从而为开发基于DEP的干细胞应用提供了基础。
The integration of microscale engineering, microfluidics, and AC electrokinetics such as dielectrophoresis has generated novel microsystems that enable quantitative analysis of cellular phenotype, function, and physiology. These systems are increasingly being used to assess diverse cell types, such as stem cells, so it becomes critical to thoroughly evaluate whether the systems themselves impact cell function. For example, engineered microsystems have been utilized to investigate neural stem/progenitor cells (NSPCs), which are of interest due to their potential to treat CNS disease and injury. Analysis by dielectrophoresis (DEP) microsystems determined that unlabeled NSPCs with distinct fate potential have previously unrecognized distinguishing electrophysiological characteristics, suggesting that NSPCs could be isolated by DEP microsystems without the use of cell type specific labels. To gauge the potential impact of DEP sorting on NSPCs, we investigated whether electric field exposure of varying times affected survival, proliferation, or fate potential of NSPCs in suspension. We found short-term DEP exposure (1 min or less) had no effect on NSPC survival, proliferation, or fate potential revealed by differentiation. Moreover, NSPC proliferation (measured by DNA synthesis and cell cycle kinetics) and fate potential were not altered by any length of DEP exposure (up to 30 min). However, lengthy exposure (> 5 min) to frequencies near the crossover frequency (50–100 kHz) led to decreased survival of NSPCs (maximum ~30% cell loss after 30 min). Based on experimental observations and mathematical simulations of cells in suspension, we find that frequencies near the crossover frequency generate an induced transmembrane potential that results in cell swelling and rupture. This is in contrast to the case for adherent cells since negative DEP frequencies lower than the crossover frequency generate the highest induced transmembrane potential and damage for these cells. We clarify contrasting effects of DEP on adherent and suspended cells, which are related to the cell position within the electric field and the strength of the electric field at specific distances from the electrodes. Modeling of electrode configurations predicts optimal designs to induce cell movement by DEP while limiting the induced transmembrane potential. We find DEP electric fields are not harmful to stem cells in suspension at short exposure times, thus providing a basis for developing DEP-based applications for stem cells.
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影响因子：
4.8
作者：
Konig, S;Hinard, V;Bernheim, L
通讯作者：
Bernheim, L
影响因子：
3.7
作者：
Labeed FH;Lu J;Mulhall HJ;Marchenko SA;Hoettges KF;Estrada LC;Lee AP;Hughes MP;Flanagan LA
通讯作者：
Flanagan LA
影响因子：
3.8
作者：
Altomare, L;Borgatti, M;Gambari, R
通讯作者：
Gambari, R
DOI：
10.1089/152581699319939
发表时间：
1999-10-01
期刊：
JOURNAL OF HEMATOTHERAPY & STEM CELL RESEARCH
影响因子：
--
作者：
Huang, Y;Yang, J;Gascoyne, PRC
通讯作者：
Gascoyne, PRC
影响因子：
7.4
作者：
Huang, Y;Joo, S;Xu, X
通讯作者：
Xu, X