Zhang Chen, Deng Yuanying, Dai Hongmei, Zhou Wenjuan, Tian Jing, Bing Guoying, Zhao Lingling
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Department of Anatomy and Neurobiology, University of Kentucky, School of Medicine, Lexington, KY 40502, USA.
Brain Res Bull. 2017 Jan;128:34-39. doi: 10.1016/j.brainresbull.2016.11.004. Epub 2016 Nov 9.
Dimethyl sulfoxide (DMSO) is a widely used solvent and vehicle for in vivo and in vitro administration of test compounds. Effects of DMSO independent of the test compound, such as in studies examining morphological plasticity or neurotoxic responses, may lead to spurious results.
To investigate effects of DMSO concentration ([DMSO]) on morphology and survival of primary cultured neurons and astrocytes.
Primary cultured neurons and astrocytes were treated with 0.25%-10.00% [DMSO] for 12-48h. Viable cell number and morphology were compared to untreated cultures using the CCK-8 assay and phase-contrast microscopy. Expression levels of the neuronal marker NeuN and astrocyte marker glial fibrillary acidic protein (GFAP) were determined by immunofluorescence and western blotting.
A [DMSO]≤0.50% had no effect on neuronal number or NeuN expression up to 24h, while ≥1.00% induced a progressive and dramatic loss of both viability and NeuN expression even after 12h. Brief (12h) exposure to ≤1.00% DMSO had no effect on astrocytes survival or GFAP expression, while ≥5.00% significantly reduced both at all exposure durations. In contrast to neurons, exposure to 0.50% and 1.00% DMSO for 24 or 48h enhanced astrocytes proliferation and GFAP expression. Astrocytic processes were maintained at 0.50% and 1.00% DMSO, while neurons exhibited marked neurite retraction at ≥0.50%.
A [DMSO]≥0.5% markedly disrupts neuronal morphology and reduces viability, even after brief exposure. In astrocytes, 0.50% and 1.00% DMSO appear to induce reactive gliosis. For treatment of neural cells, [DMSO] should be ≤0.25% to obviate spurious vehicle effects.
二甲基亚砜（DMSO）是一种广泛用于体内和体外给予受试化合物的溶剂和载体。在研究形态可塑性或神经毒性反应等实验中，DMSO独立于受试化合物产生的效应可能会导致虚假结果。
研究DMSO浓度（[DMSO]）对原代培养神经元和星形胶质细胞形态及存活的影响。
用0.25%-10.00%的[DMSO]处理原代培养的神经元和星形胶质细胞12-48小时。使用CCK-8法和相差显微镜将活细胞数量和形态与未处理的培养物进行比较。通过免疫荧光和蛋白质印迹法测定神经元标志物NeuN和星形胶质细胞标志物胶质纤维酸性蛋白（GFAP）的表达水平。
[DMSO]≤0.50%在长达24小时内对神经元数量或NeuN表达无影响，而≥1.00%即使在12小时后也会导致活力和NeuN表达逐渐且显著丧失。短暂（12小时）暴露于≤1.00%的DMSO对星形胶质细胞存活或GFAP表达无影响，而≥5.00%在所有暴露时长下均显著降低两者。与神经元不同，暴露于0.50%和1.00%的DMSO 24或48小时可增强星形胶质细胞增殖和GFAP表达。在0.50%和1.00%的DMSO浓度下，星形胶质细胞的突起得以维持，而神经元在≥0.50%时则表现出明显的神经突回缩。
即使短暂暴露，[DMSO]≥0.5%也会显著破坏神经元形态并降低活力。在星形胶质细胞中，0.50%和1.00%的DMSO似乎会诱导反应性胶质增生。对于神经细胞的处理，[DMSO]应≤0.25%以避免虚假的载体效应。
