Umbilical cord mesenchymal stem cells undergoing hypoxic preconditioning can differentiate into vascular endothelial-like cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Incidence of diabetic lower extremity vascular disease is increasing, so how to improve blood vessels of the lower limbs and increase angiogenesis becomes the research focus. Umbilical cord mesenchymal stem cells have been employed clinical y via local intramuscular injection, but the specific therapeutic effect and mechanism are not clear. OBJECTIVE:To investigate the effects of hypoxic preconditioning and cobalt chloride medium on the differentiation of umbilical cord mesenchymal stem cells into vascular endothelial-like cells in vitro. METHODS:Umbilical cord mesenchymal stem cells were isolated and cultured, and then treated with different concentrations of cobalt chloride. Enzyme linked immunosorbent assay was used to detect levels of basic fibroblast growth factor and vascular endothelial growth factor gene in cellsupernatants, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to detect cellproliferation. The safety of umbilical cord mesenchymal stem cells before and after cobalt chloride induction was detected using chromosome. The umbilical cord mesenchymal stem cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10μg/L vascular endothelial growth factor, 10μg/L basic fibroblast growth factor and 10%fetal bovine serum, which were induced to differentiate into vascular endothelial-like cells. Endothelial-like cellphenotype CD31 and von Wil ebrand factor were identified before and after induction, through the observation of three-dimensional model of angiogenesis, the angiogenic capacity of umbilical cord mesenchymal stem cells was determined. RESULTS AND CONCLUSION:Umbilical cord mesenchymal stem cells strongly expressed the surface marks. After the cobalt chloride induction, the proliferation of umbilical cord mesenchymal stem cells was positivelycorrelated with the time of induction. Based on the levels of vascular endothelial growth factor and basic fibroblast growth factor, the optimal concentration of cobalt chloride was 200 μmol/L. Chromosome detection showed the stability of cells after cobalt chloride induction. After induction, CD31 and von Wil ebrand factor were strongly expressed. Three-dimensional observation showed umbilical cord mesenchymal stem cells could be induced to form the lumen-like structure with different diameter sizes, and umbilical cord mesenchymal stem cells could be induced to differentiate into endothelial-like cells, and have a angiogenic capacity. RESULTS AND CONCLUSION:Umbilical cord mesenchymal stem cells strongly expressed the surface marks. After the cobalt chloride induction, the proliferation of umbilical cord mesenchymal stem cells was positivelycorrelated with the time of induction. Based on the levels of vascular endothelial growth factor and basic fibroblast growth factor, the optimal concentration of cobalt chloride was 200 μmol/L. Chromosome detection showed the stability of cells after cobalt chloride induction. After induction, CD31 and von Wil ebrand factor were strongly expressed. Three-dimensional observation showed umbilical cord mesenchymal stem cells could be induced to form the lumen-like structure with different diameter sizes, and umbilical cord mesenchymal stem cells could be induced to differentiate into endothelial-like cells, and have a angiogenic capacity.