锌离子促进人骨髓间充质干细胞增殖并增强其纤黏连蛋白表达的实验研究

Zinc Ion Promotes the Proliferation and Fibronectin Expression of Human Bone Marrow Mesenchymal Stem Cells

目的探索适宜的锌离子浓度,以促进人间充质干细胞(Mesenchymal stem cells,MSC)在无血清培养基中的黏附和增殖。方法将人骨髓MSC培养于含胰岛素、转铁蛋白和亚硒酸钠的α-MEM中,体系中加入不同浓度硫酸锌。培养72 h后,MTT实验测定490 nm光密度值,观察细胞增殖状态。蛋白印迹杂交比较含锌离子(1×10-5M)培养体系与未加锌离子培养体系两组细胞纤黏连蛋白含量的差异。结果MTT结果显示,在一定浓度范围内,锌离子促进人骨髓MSC增殖,作用呈浓度依赖性,适宜浓度为1×10-5M;蛋白印迹实验表明,在该浓度刺激下细胞培养72 h后,纤黏连蛋白表达量显著升高。结论锌离子可作为人无血清培养基中的一个成分,可提高人骨髓MSC的贴壁生长能力。

Objective To investigate the appropriate concentration of zinc ion in the serum-free medium promoting the attachment and proliferation of human mesenchymal stem cells （MSCs）. Methods Human bone marrow MSCs were maintained in α-MEM containing insulin, transferrin and selenite sodium, with or without graded concentrations of zinc ion. Optical density at 490 nm was detected with MTT assay to evaluate the proliferation status of MSCs cultured for 72 hours. MSCs were cultured in the presence of zinc ion （1×10^5M） for 72 hours, and the cellular content of fibronectin was detected by western blotting and compared with that of MSCs cultured without zinc ion. Results MTr assay showed that zinc ion within a range of concentrations promoted human MSC growth in a dose-dependent manner. MSCs cultured with zinc ion （1×10^55 M） for 72 hours was significantly higher than that of the counterparts without zinc ion. Furthermore, after stimulation in the presence of 1×10^55 M zinc ion for 72 hours, the cellular content of fibronectin was greatly higher than that of MSCs in the other concentrations of zinc ion. Conclusion Zinc ion could be served as one of the compositions in the serum-free medium to promote the attachment and proliferation of human bone marrow MSCs.