Quantitative study of electroporation-mediated molecular uptake and cell viability
Quantitative study of electroporation-mediated molecular uptake and cell viability
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DOI：
10.1016/s0006-3495(01)76055-9
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发表时间：
2001-02-01
影响因子：
3.4
通讯作者：
Prausnitz, MR
中科院分区：
文献类型：
--
作者：
Canatella, PJ;Karr, JF;Prausnitz, MR
电穿孔在实验室转染和临床化疗中的应用受到对电穿孔参数对分子摄取和细胞活力的影响的不完全理解的限制。为了解决这一需求，使用流式细胞术对超过200种不同的实验条件组合定量DU 145前列腺癌细胞的钙黄绿素摄取和活力。实验参数包括场强（0.1- 3.3kV/cm）、脉冲长度（0.05-20 ms）、脉冲数（1-10）、钙黄绿素浓度（10-100 μ M）和细胞浓度（0.6-23%体积）。这些数据表明，无论是电荷还是能量都不是电穿孔效果的良好预测因子。相反，摄取和活力表现出复杂的依赖于场强，脉冲长度和脉冲数。细胞浓度的影响被定量解释由相邻细胞引起的电场扰动。摄取量与外部钙黄绿素浓度呈线性关系。这个大的定量数据集可用于优化电穿孔方案，测试理论模型，并指导机理解释。
Electroporation's use for laboratory transfection and clinical chemotherapy is limited by an incomplete understanding of the effects of electroporation parameters on molecular uptake and cell viability. To address this need, uptake of calcein and viability of DU 145 prostate cancer cells were quantified using flow cytometry for more than 200 different combinations of experimental conditions. The experimental parameters included field strength (0.1-3.3 kV/cm), pulse length (0.05-20 ms), number of pulses (1-10), calcein concentration (10-100 muM), and cell concentration (0.6-23% by volume). These data indicate that neither electrical charge nor energy was a good predictor of electroporation's effects. Instead, both uptake and viability showed a complex dependence on field strength, pulse length, and number of pulses. The effect of cell concentration was explained quantitatively by electric field perturbations caused by neighboring cells. Uptake was shown to vary linearly with external calcein concentration. This large quantitative data set may be used to optimize electroporation protocols, test theoretical models, and guide mechanistic interpretations.