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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 HPMSCS transfected with pIRES2-EGFPhVEGF165 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.