The assessment of electrophysiological activity in human-induced pluripotent stem cell-derived cardiomyocytes exposed to dimethyl sulfoxide and ethanol by manual patch clamp and multi-electrode array system
The assessment of electrophysiological activity in human-induced pluripotent stem cell-derived cardiomyocytes exposed to dimethyl sulfoxide and ethanol by manual patch clamp and multi-electrode array system
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DOI：
10.1016/j.vascn.2017.03.003
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发表时间：
2017-09-01
影响因子：
1.9
通讯作者：
Seo, Joung-Wook
中科院分区：
文献类型：
--
作者：
Hyun, Soo-Wang;Kim, Bo-Ram;Seo, Joung-Wook
简介：最近，电生理活性已被有效地测量在人类诱导的多能干细胞衍生的心肌细胞（hiPSC-CM），以预测药物诱导的心律失常。在许多实验中，二甲基亚砜（DMSO）和乙醇被用作稀释剂。然而，在基于膜片钳和多电极阵列（MEA）的hiPSC-CMs中，可以有效地用于电生理参数测量的最大DMSO和乙醇浓度尚未完全阐明。方法：我们使用膜片钳和MEA研究了用作稀释剂的不同浓度的DMSO和乙醇对hiPSC-CMs中几个电生理参数的影响。外部溶液中浓度> 1%的DMSO和乙醇均导致摩尔渗透压浓度>400 mOsmol/kg，但pH不受任一试剂的影响。在检测浓度下，DMSO和乙醇均未导致细胞死亡。而在1%乙醇浓度下，静息膜电位、动作电位振幅、90%和40%时的动作电位时程和校正场电位时程均显著降低。1%的DMSO也显著降低了钠峰幅度。此外，在3%的DMSO和乙醇浓度下，动作电位和场电位的波形都记录为不规则的。浓度高达0.3%的任何一个代理不影响渗透压，pH值，细胞死亡，或在hiPSC-CMs.Discussion电生理参数：我们的研究结果表明，0.3%是最大浓度，DMSO或乙醇应用于稀释目的的hiPSC-CMs为基础的膜片钳和MEA。
Introduction: Recently, electrophysiological activity has been effectively measured in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to predict drug-induced arrhythmia. Dimethyl sulfoxide (DMSO) and ethanol have been used as diluting agents in many experiments. However, the maximum DMSO and ethanol concentrations that can be effectively used in the measurement of electrophysiological parameters in hiPSC-CMs-based patch clamp and multi-electrode array (MEA) have not been fully elucidated.Methods: We investigated the effects of varying concentrations of DMSO and ethanol used as diluting agents on several electrophysiological parameters in hiPSC-CMs using patch clamp and MEA.Results: Both DMSO and ethanol at concentrations > 1% in external solution resulted in osmolality >400 mOsmol/kg, but pH was not affected by either agent. Neither DMSO nor ethanol led to cell death at the concentrations examined. However, resting membrane potential, action potential amplitude, action potential duration at 90% and 40%, and corrected field potential duration were decreased significantly at 1% ethanol concentration. DMSO at 1% also significantly decreased the sodium spike amplitude. In addition, the waveform of action potential and field potential was recorded as irregular at 3% concentrations of both DMSO and ethanol. Concentrations of up to 0.3% of either agent did not affect osmolality, pH, cell death, or electrophysiological parameters in hiPSC-CMs.Discussion: Our findings suggest that 0.3% is the maximum concentration at which DMSO or ethanol should be used for dilution purposes in hiPSC-CMs-based patch clamp and MEA.