TY - JOUR
T1 - Effects of dimethyl sulfoxide on the morphology and viability of primary cultured neurons and astrocytes
AU - Zhang, Chen
AU - Deng, Yuanying
AU - Dai, Hongmei
AU - Zhou, Wenjuan
AU - Tian, Jing
AU - Bing, Guoying
AU - Zhao, Lingling
N1 - Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - Background 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. Aim To investigate effects of DMSO concentration ([DMSO]) on morphology and survival of primary cultured neurons and astrocytes. Methods Primary cultured neurons and astrocytes were treated with 0.25%–10.00% [DMSO] for 12–48 h. 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. Results A [DMSO] ≤ 0.50% had no effect on neuronal number or NeuN expression up to 24 h, while ≥1.00% induced a progressive and dramatic loss of both viability and NeuN expression even after 12 h. Brief (12 h) 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 48 h 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%. Conclusion 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.
AB - Background 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. Aim To investigate effects of DMSO concentration ([DMSO]) on morphology and survival of primary cultured neurons and astrocytes. Methods Primary cultured neurons and astrocytes were treated with 0.25%–10.00% [DMSO] for 12–48 h. 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. Results A [DMSO] ≤ 0.50% had no effect on neuronal number or NeuN expression up to 24 h, while ≥1.00% induced a progressive and dramatic loss of both viability and NeuN expression even after 12 h. Brief (12 h) 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 48 h 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%. Conclusion 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.
KW - Astrocytes
KW - Dimethyl sulfoxide
KW - Morphology
KW - Neurons
KW - Primary culture
KW - Survival
UR - http://www.scopus.com/inward/record.url?scp=84997447914&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84997447914&partnerID=8YFLogxK
U2 - 10.1016/j.brainresbull.2016.11.004
DO - 10.1016/j.brainresbull.2016.11.004
M3 - Article
C2 - 27836802
AN - SCOPUS:84997447914
SN - 0361-9230
VL - 128
SP - 34
EP - 39
JO - Brain Research Bulletin
JF - Brain Research Bulletin
ER -