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[The establishment and validation of an endothelial cell senescence model induced by carbamide peroxide]
Quan-bo Zhang1, Bi-hua Wu, Yu-feng Qing
1Department of Geriatric, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
Objective:
To establish an in vitro pig iliac artery endothelial cells (PIECs) senescence model using carbamide peroxide (CP).
Methods:
MTT assay and DAPI staining were used to define the optimal concentration of CP for inducing to the PIECs senescence model. Cellular morphology, MTT assay, EdU labeling, SA-β-gal staining and cell scratch test were performed to analyze the cell growth kinetic, proliferative activity, aging ratio and migratory activity difference post CP induction. PI signal staining flow cytometry was used to analyze the cell cycle distribution difference of cells before and after CP induction.
Results:
The optimal CP concentration was 40 µmol/L to induce PIECs senescence. After 1 h treatment with 40 µmol/L CP, the PIECs presented typical aging form with lager and more rounded shapes. Compared with control group, the proliferative activity and the migratory distance of CP group were significantly decreased; the SA-β-gal staining positive ratio was significantly increased; the data of mitotic cycle distribution with flow cytometry analysis showed that most cells were arrested at G(1)/G(0) phase.
Conclusion:
CP could efficiently induce pig iliac artery endothelial cell senescence in vitro.
