Effects of dimethyl sulfoxide on the morphology and viability of primary cultured neurons and astrocytes
Abstract
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 12-48h. 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.Results 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%.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.
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The final DMSO concentration did not exceed 0.15% in any of the recordings.
“The final DMSO concentration did not exceed 0.15% in any of the recordings given DMSO effects at higher concentrations on neuronal viability or interference with synaptic transmission (Nakahiro et al., 1992;Tsvyetlynska et al., 2005;Zhang et al., 2017).”
From Alcohol dependence and withdrawal increase sensitivity of central amygdalar GABAergic synapses to the glucocorticoid receptor antagonist mifepristone in male rats (2022) · cites this paper as Zhang et al., 2017
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- Authors
- Zhang, C., Deng, Y., Dai, H., Zhou, W., Tian, J., Bing, G., & Zhao, L.
- Affiliations
- Central South University (CN)
- University of Kentucky (US)
- Corresponding author
- Zhao, Lingling
- Venue
- Brain Research Bulletin
- Publisher
- Elsevier BV
- Published
- 2017-01
- Volume
- 128
- Pages
- 34-39
- ISSN
- 0361-9230 (ISSN-L)
- OpenAlex
- W2556629357
- Cited by
- 62
- References
- 29
- Citations by year
- 2026: 7 · 2025: 15 · 2024: 8 · 2023: 4 · 2022: 3 · 2021: 5 · 2020: 11 · 2019: 5 · 2018: 4
- FWCI
- 4.61
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Brain Research Bulletin (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Third Xiangya Hospital of Central South University
- Hunan Province
- Topics
- Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Immunology and Microbiology › Immunology › Immunotoxicology and immune responses
- Keywords
- dimethyl sulfoxide, vehicle effects, primary neuronal cultures, astrocytes, neuronal viability, cell morphology, neurite retraction, reactive gliosis, glial fibrillary acidic protein, NeuN, astrocyte proliferation, DMSO concentration
- MeSH terms
- Animals, Astrocytes, Cell Survival, Cells, Cultured, Cerebral Cortex, Dimethyl Sulfoxide, Dose-Response Relationship, Drug, Glial Fibrillary Acidic Protein, Nerve Tissue Proteins, Neurons, Time Factors, Rats, Sprague-Dawley, Antigens, Nuclear
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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