目的:探讨转录因子SOX2对牙髓干细胞(DPSCs)神经分化的影响,为研究DPSCs神经分化的机制提供依据。方法:从人源第三磨牙牙髓中分离DPSCs(DPSCs组),通过逆转录病毒感染方法构建过表达SOX2的DPSCs(DPSCs-SOX2组),以空白载体感染的DPSCs作为对照组(DPSCs-vector组),采用神经分化培养基诱导各组DPSCs分化为神经前体细胞(NPCs)。采用CCK-8法分析各组NPCs的增殖指数(PI);采用qPCR和流式细胞术检测各组NPCs中Nestin和Pax6基因的表达水平;采用免疫荧光和流式细胞术分析各组NPCs在神经元分化方面的能力和分化后β3-Tubulin的表达效率。结果:通过神经分化培养基的定向诱导,正常DPSCs、DPSCs-vector和DPSCs-SOX2均可通过分化形成NPCs,但DPSCs-SOX2组NPCs的PI和NPCs中Nestin和Pax6表达水平明显高于DPSCs组和DPSCs-vector组(P〈0.05);DPSCs-SOX2组NPCs在神经元分化方面也具有一定的优势,分化后细胞的β3-Tubulin表达效率明显高于其他2组(P〈0.05)。结论:SOX2对DPSCs的神经分化具有一定的促进作用。
Objective:To explore the effect of SOX2 on the neural differentiation of dental pulp stem cells(DPSCs),and to provide basis for studying the mechanism of neural differentiation of DPSCs.Methods:The DPSCs(DPSCs group)were isolated from human dental pulp of third molar,and the SOX2 over exression DPSCs(DPSCs-SOX2group)were established via retrovirus infection.The empty vector-infected DPSCs(DPSCs-vector group)were regarded as control group.The DPSCs in various groups were induced into neural progenitor cells(NPCs)by neural differentiation medium.The proliferation index(PI)of NPCs in various groups was evaluated by CCK-8method;the expression levels of Nestin and Pax6 gene in NPCs in various groups were detected by qPCR and FACS;the differentiation abilities of NPCs and the expression efficiencies ofβ3-Tubulin after differentiation in various groups were evaluated by immunofluorescence and FACS methods.Results:All of normal DPSCs,DPSCsvector and DPSCs-SOX2 derived from NPCs could be differentiated into NPCs successfully through the induction of neural differentiation medium.Compared with DPSCs and DPSCs-vector groups,the PI of DPSCs in DPSCs-SOX2 group was increased(P〈0.05),and the expression levels of Nestin and Pax6 in NPCs were increased(P〈0.05).The expression efficiency ofβ3-Tubulin in differentiated NPCs in DPSCs-SOX2 group was higher than those in DPSCs and DPSCs-vector groups(P〈0.05).Conclusion:SOX2could have a certain promotive effect on the neural differentiation of DPSCs.