Establishment of anoxia model of human dental pulp stem cells induced by cobalt chloride in vitro
Objective:To observe the effect of cobalt chloride(CoCl2)on primary human dental pulp stem cells in vitro,and to explore a chemical intervention model for hypoxia in vitro.Methods:Primary human dental pulp stem cells were cultured by adherent method and treated with 50,100,200,300 μmol/L CoCl2 solution for 48 h,respectively.The cell growth and apoptosis were observed by CCK-8 assay under different conditions.The expressions of hypoxia-inducing factor 1α(HIF-1α)were assessed and quantitative real-time PCR.Results:Compared with blank control group,the dental pulp stem cells overall trend of cell survival rate in CoCl2 treatment group was significantly reduced with the increase of concentration(P<0.05),but the survival rate of was significantly increased under the induction of low concentration of CoCl2.The mRNA lev-els of HIF-1α were also significantly increased with the increase of CoCl2 concentration(P<0.05).However,the expression of HIF-1α reached a plateau under the induction of 100 μmol/L CoCl2.Conclusion:The proliferation,apoptosis and expression of related genes in dental pulp stem cells are correlated with the concentration of CoCl2,showing an overall increasing trend.This study successfully established an in vitro culture method for inducing hypoxia in dental pulp stem cells which can serve as a reference for studying the hypoxia model of dental pulp stem cells.
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