Vitamin C maintaining bone marrow mesenchymal stem cell self-renewal

LI Xiao-hong; ZHU he-ning; JIANG Xue-yan; DENG Chun-yu; KUANG Su-juan; ZHANG Guang-feng; FU Yong-heng; YU Xi-yong

摘要：BackgroundBone marrow mesenchymal stem cells(BMSCs)can be isolated and cultured to many passages.However, Stem cells including BMSCs quickly undergo senescence in culture.The cell senescence and multidirectional differentiation have hampered producing BMSCs in quantity with their undifferentiated state. In this study we report a natural compound,vitamin C(Vc),maintains BMSCs stem property.Methods Human BMSCs were isolated from bone marrow and purified by 1.073 g/mL density gradient centrifugation.50 ng/mL Vc were added to BMSCs for different time point.Flowcytometry was used to detect cell surface markers of BMSCs with or without Vc treatment. BMSCs proliferation was analyzed by MTT assay.PCR(Polymerase chain reaction)and real-time PCR were used for detecting c—kit,nanog,and Oct-4 genes expression levels.DNA methyltransferase(Dnmt)1 and Dnmt3b levels were also detected by real-time PCR.Results Flowcytometry showed that after Vc treatment for 6 h,the surface markers of BMSCs were almost unchanged.Vc increased the proliferation activity of BMSCs from 6h to 24 h.PCR showed the expression of c-kit,nanog,and oct一4 genes were obviously increased in Vc treated group than control group at 12 h.Real-time PCR showed that the level of c-kit,nanog,and oct-4 genes were unregulated from 6h to 12h compared with control group.Vc also increased Dnmt3b but not Dnmt1 gene expression.Conclusions Our results showed Vc acts at least accelerates BMSCs proliferation and maintains stem cell property.In our study,we highlighted a method of improving the speed of BMSCs generation and provided additional insights into the mechanistic basis of preventing BMSCs senescence.