氯化锂对牙髓干细胞增殖与分化的影响

Effects of lithium chloride on proliferation and differentiation of human dental pulp stem cells

关键词:  氯化锂  牙髓干细胞  细胞增殖  细胞分化

Key words:  lithium chloride  dental pulp stem cell  cell proliferation  cell differentiation

摘要:

目的 检测氯化锂对人牙髓干细胞（dental pulp stem cells， DPSCs)细胞增殖与分化的影响。方法 收集18~25岁患者因正畸拔除的健康完整前磨牙或智齿，用改良组织块法分离培养牙髓干细胞；利用免疫荧光染色与流式细胞术对分离培养的细胞进行细胞来源鉴定；利用细胞计数试剂盒（CCK-8）测定不同浓度氯化锂对牙髓干细胞增殖的影响，选择适宜浓度的氯化锂对牙髓干细胞进行矿化诱导培养，利用茜素红染色观察矿化结节形成以及荧光定量PCR实验研究氯化锂对牙髓干细胞分化的影响。结果 流式细胞术及免疫组化染色结果显示原代人牙髓干细胞来源与标志物与间充质干细胞表现一致；CCK-8法检测结果表明2.5mmol/L的氯化锂对牙髓干细胞增殖具有促进作用；选用该浓度下的氯化锂对牙髓干细胞进行矿化诱导培养，茜素红染色与荧光定量PCR结果显示，氯化锂对牙髓干细胞牙向分化具有促进作用。结论 一定浓度的氯化锂对牙髓干细胞细胞增殖与细胞分化具有促进作用。

Abstract:

Objective To investigate the effect of lithium chloride on the proliferation and differentiation of human dental pulp stem cells. Methods The healthy premolars or third molars extracted due to orthodontics were collected from 18 to 25-year-old patients. The dental pulp stem cells were isolated and cultured by modified tissue block method. Immunofluorescence staining and flow cytometry were used to identify the origin of cells. The cell count kit（CCK-8) was used to detect the effect of different concentrations of lithium chloride on the proliferation of dental pulp stem cells. The appropriate concentration of lithium chloride was selected to induce the mineralization of the dental pulp stem cells. The mineralized nodule was observed by alizarin red staining, and the real-time PCR were used to detect the effect of lithium chloride on the differentiation of dental pulp stem cells. Results Flow cytometry and immunohistochemical staining showed that the source and markers of the primary human dental pulp stem cells were consistent with mesenchymal stem cells; the result of CCK-8 indicated that with a concentration of 2.5mmol/L, lithium chloride promoted the proliferation of dental pulp stem cells. Then the cells were cultured with 2.5mmol/L lithium chloride to induce osteogenic differentiation and the results of alizarin red staining and qPCR showed that lithium chloride promoted the dentinogenic and osteogenic differentiation of dental pulp stem cells. Conclusion A certain concentration of lithium chloride might promote the proliferation and differen-tiation of dental pulp stem cells.