山东医药2017，Vol.57Issue(32)：25-28,4.DOI:10.3969/j.issn.1002-266X.2017.32.007
低氧预处理对脐带间充质干细胞表型、增殖及神经营养因子分泌功能的影响
Effects of hypoxic preconditioning on cell phenotype, proliferation, and secretion of neurotrophic factors of umbilical cord mesenchymal stem cells
摘要
Abstract
Objective To investigate the influences of hypoxic preconditioning on cell phenotype, proliferation, and secretion of neurotrophic factors of human umbilical cord mesenchymal stem cells (UCMSCs).Methods The discarded umbilical cord tissues during delivery process were collected for UCMSCs culture, and the primary cultured mesenchymal stem cells were passaged.Cells at passage 3 were randomly divided into the normoxia group and hypoxia group, which were growing in 21% and 5% O2, respectively.Cells in logarithmic growth phase were used.The expression of CD44, CD45, CD90, and HLA-DR was detected by flow cytometry;the expression of CD34, laminin, and vimentin was determined by immunocytochemistry;the mRNA expression of brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and ciliary neurotrophic factor (CNTF) was determined by real-timer fluorescent quantitative PCR.After the cells were cultured for 1 to 6 days, we did the cell counting, and determined the cell proliferation by CCK-8.Results CD44, CD90, laminin, and vimentin were highly expressed, and CD45, HLA-DR, and CD34 were low expressed in both of the two groups, which showed that cells in the two groups had the characteristics of mesenchymal stem cells.There was no significant difference in expression of phenotype-related indexes between the two groups (all P>0.05).The CCK-8 results indicated that the cell counting and proliferation rate first increased and then decreased in 1-6 days′ culture in the two groups.When the cells were cultured for 1-6 days, the cell counting in the hypoxia group was higher than that in the normoxia group;when the cells were cultured for 2-6 days, the proliferation activity of the hypoxia group was higher than that of the normoxia group (all P<0.05).The mRNA expression of HGF, BDNF, VEGF, and CNTF in the hypoxia group was higher than that of the normoxia group (all P<0.05).Conclusion Hypoxic preconditioning has little effect on the phenotypic features of UCMSCs, but can significantly enhance the proliferation rate and the secretion of neurotrophic factors.关键词
间充质干细胞/脐带/低氧/细胞增殖/细胞表型/神经营养因子Key words
mesenchymal stem cells/umbilical cord/hypoxia/cell proliferation/cell phenotype/neurotrophic factors分类
医药卫生引用本文复制引用
姚舜,靳安民,张辉,闵少雄,黄帅,周治来..低氧预处理对脐带间充质干细胞表型、增殖及神经营养因子分泌功能的影响[J].山东医药,2017,57(32):25-28,4.基金项目
国家自然科学基金青年科学基金资助项目(81400996) （81400996）
广东省自然科学基金博士启动项目(2014A030310100). （2014A030310100）
