Cerebral infarction causes severe morbidity and mortality. Most patients with cerebral infarction should take antiplatelet drugs daily, so the effects of those drugs on the regeneration of the brain need to be investigated. Aspirin and clopidogrel are the most widely used antiplatelet drugs for the prevention of ischemic stroke. We investigated the effects of aspirin and clopidogrel on neural stem cells (NSCs). NSCs were dissociated from fetal rat cortex and cultured with basic fibroblast growth factor and N2 medium. To measure the effects of aspirin and clopidogrel on NSCs, NSCs were treated with several concentrations of aspirin, clopidogrel bisulfate, and clopidogrel resinate for 24 h. After the treatment, we measured cell viability by cell counting kit-8, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, trypan blue staining, flow cytometry, and lactate dehydrogenase assay. To evaluate their effects on NSC proliferation, we performed BrdU cell proliferation assay and colony-forming unit assay. We compared the intracellular protein level in the NSCs treated with aspirin and two types of clopidogrel, by proteomics analysis. Various viability tests showed that clopidogrel resinate and clopidogrel bisulfate did not affect the viability and proliferation of NSCs whereas aspirin decreased them even at low concentrations which are clinically relevant. Moreover, through the proteomics, it was confirmed that the toxicity of aspirin to NSCs might be associated with the alteration of several intracellular proteins. Taken together, these results suggest that clopidogrel resinate and clopidogrel bisulfate are safe but aspirin could be toxic to NSCs. Therefore, when these antiplatelet agents are prescribed over the long-term, the finding that aspirin could be toxic to NSCs should be considered.
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Neural stem cells
Central nervous system
Dual antiplatelet therapy
Basic fibroblast growth factor
Cell counting kit-8
Two-dimensional gel electrophoresis
Coomassie brilliant blue R-350
Peptide mass fingerprinting
Matrix-assisted laser desorption/ionization-time of flight mass spectrometer
NADH dehydrogenase 1 subunit 10
Creatine kinase brain-type
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This work was supported by the Basic Science Research Program of the National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning (2015R1A2A2A04004865), by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HI17C2160), and by Korea Drug Development Fund (KDDF) funded by Ministry of Science and ICT, Ministry of Trade, Industry & Energy and Ministry of Health & Welfare (KDDF-201609-02, Republic of Korea).
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Hwang, M., Park, H., Choi, H. et al. Effects of aspirin and clopidogrel on neural stem cells. Cell Biol Toxicol 34, 219–232 (2018). https://doi.org/10.1007/s10565-017-9412-y
- Neural stem cells
- Clopidogrel bisulfate
- Clopidogrel resinate