Effects of dimethyl sulfoxide on the morphology and viability of primary cultured neurons and astrocytes.
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.
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Chen
C
Zhang
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Yuanying
Y
Deng
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Hongmei
H
Dai
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Wenjuan
W
Zhou
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
Jing
J
Tian
Department of Anatomy and Neurobiology, University of Kentucky, School of Medicine, Lexington, KY 40502, USA.
Guoying
G
Bing
Department of Paediatrics, Third Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: llzhao2011@qq.com.
Lingling
L
Zhao
D000818:Animals; D034961:Antigens, Nuclear / Q000378:metabolism; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004121:Dimethyl Sulfoxide / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D034961:Antigens, Nuclear; C000614142:Gfap protein, rat; D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; C546339:Rbfox3 protein, rat; D004121:Dimethyl Sulfoxide
Astrocytes; Dimethyl sulfoxide; Morphology; Neurons; Primary culture; Survival