Abstract
The experiment adopting reverse transcription polymerase chain reaction (RT-PCR) technology, amplified hVEGF165 gene fragments from human leukemia cells HL-60. hVEGF165 gene was reconstructed in pIRES2-EGFP and transferred into the human placenta-derived mesenchymal stem cells (HPMSCs) by liposome-mediated method successfully. The mRNA and protein of hVEGF165 in the transferred cells was detected by RT-PCR and Western blot, and the results showed that hVEGF165mRNA and the protein expressed by HPMSCs transfected with pIRES2-EGFP-hVEGF165 was significantly more than HPMSCs transfected with pIRES2-EGFP. EGFP expression was observed under fluorescence microscope, which proved that the report gene was successfully transferred to the target cells. hVEGF biology activity and cell proliferation activity of HPMSCStransfected with pIRES2-EGFP-hVEGF165 was detected by MTT array, which showed that hVEGF165 can promote the proliferation of HPMSCS; however, hVEGF165 biology activity of HPMSCS transfected with pIRES2-EGFP-hVEGF165 was significantly more than HPMSCs transfected with pIRES2-EGFP. Identification of multipotentiality showed that HPMSCS transfected with pIRES2-EGFP-VEGF165 still maintained multipotentiality. Key words: Transfect, human placenta-derived mesenchymal stem cells, hVEGF165.
Highlights
Vascular endothelial growth factor is a potent factor for promoting angiogenesis, and this can induce angiogenesis (Parati et al, 2003), increase vascular permeability (Jackson et al, 2001), maintain vascular function and play an important role in wound healing
Reverse transcription polymerase chain reaction (RT-Polymerase chain reaction (PCR)) products were obtained for electrophoresis on agarose gel, and target strips appeared on about 600 bp of the DNA marker (Figure 1), which indicated that the amplification of the target gene fragment succeeded
Group A generated a positive cell clone 15 days after screening by G418 selective medium, which indicated that both effects of human placenta-derived mesenchymal stem cells (HPMSCs) transfected with pIRES2-EGFP-hVEGF165 and G418 selection are better (Figure 4). 2
Summary
Vascular endothelial growth factor is a potent factor for promoting angiogenesis, and this can induce angiogenesis (Parati et al, 2003), increase vascular permeability (Jackson et al, 2001), maintain vascular function and play an important role in wound healing. It was discovered that the bone marrow mesenchymal stem cells (Halleux et al, 2001) and adipose-derived stem cells (Lei et al, 2007) had important application potential in the field of wound healing in recent years, in that they can locally extend homing to trauma (Kajstura et al, 2005) and directionally differentiate them into wound repair cells, endothelial cells and fibroblasts (Brittan et al, 2005).
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