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- W3029958715 abstract "ObjectiveTo build the lentiviral vectors of pigment epithelial derived factor (PEDF) gene, and investigate their expression in human umbilical cord mesenchymal stem cells (hUCMSCs).MethodsThe PEDF lentiviral vectors (LV-PEDF) were built by DNA recombination and confirmed by DNA sequencing. hUCMSCs were transfected by LV-PEDF with MOI 10, 30, 50, respectively. The transfection efficiency was observed under fluorescence microscope. Cell immunofluorescence, immunocytochemistry and real-time PCR methods were used for detecting the expression of PEDF and VEGF.ResultsThe PEDF cDNA was sub-cloned into pCDH-CMV-MCS-EF1-copGFP vector successfully. DNA sequencing analysis confirmed that PEDF gene sequence was exactly the same with that reported in GenBank. pCDH-PEDF infected cells could show green fluorescence under fluorescence microscope. The transfection efficiency was 72.1% in PEDF-MSCs. Immunofluorescence and immunochemical staining confirmed that PEDF protein was overexpressed in hUCMSCs. The relative expression of PEDF mRNA in experimental group and control group was(0.170±0.028) and (0.015±0.007) respectively by RT-PCR, the difference was statistically significant(P>0.001). The relative expression levels of VEGF mRNA in the two groups were (0.265±0.022) and(0.285±0.049), respectively, with no significant difference (P>0.05).ConclusionsWe successfully built a lentivirus vector carrying PEDF gene and obtained hUCMSCs with overexpressed PEDF.Key words: Vascular endothelial growth factors; Mesenchymal stem cells; Lentivirus infections; Cells, cultured" @default.
- W3029958715 created "2020-06-05" @default.
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- W3029958715 date "2017-11-25" @default.
- W3029958715 modified "2023-09-24" @default.
- W3029958715 title "Lentiviral transfection of pigment epithelial derived factor gene into human umbilical cord mesenchymal stem cells" @default.
- W3029958715 doi "https://doi.org/10.3760/cma.j.issn.1005-1015.2017.06.016" @default.
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