Effects of hypoxic preconditioning on cell phenotype, proliferation, and secretion of neurotrophic factors of umbilical cord mesenchymal stem cells
YAO Shun
JIN Anmin
ZHANG Hui
MIN Shaoxiong
HUANG Shuai
ZHOU Zhilai
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.
Keywords：mesenchymal stem cellsumbilical cordhypoxiacell proliferationcell phenotypeneurotrophic factors
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 25-28 )