Non-viral vector-load enhanced green fluorescent protein transfection of immortalized neural precursor cells.doc

 PAGE \* MERGEFORMAT 8 Non-viral vector-load enhanced green fluorescent protein transfection of immortalized neural precursor cells Authors: Gui clever, Zhiheng, Zhang Chuan Han, the peak [Keywords:] Green fluorescent protein;, gene expression;, transfection;, neural precursor cells Transfection of enhanced green fluorescent protein into immortalized neural progenitor cells by nonviral vectors [Abstract] AIM: To investigate an effective transfection method of nonviral vectors and the expression of enhanced green fluorescent protein (EGFP) in the transfected immortalized neural progenitor cells (INPC). METHODS: The plasmid EGFPC1 was transfected into INPC by three different nonviral vectors respectively, including Lipofectamine 2000, TRANSfection and Sofast. The expression of EGFP and the transfection efficiency of the 3 vectors were measured at 24 h after transfection. The transfection efficiency of the vector which was the most efficient, was detected 12, 24, 48 and 72 h after transfection to determine the expression peak of EGFP. The viability of transfected and nontransfected cells was measured by trypan blue rejection test. After the plasmid EGFPC1 was transfected into INPC, the stable cell clone designated INPC / EGFP was isolated by G418 selection. The positive rate of EGFP was observed. And the specific molecular marker of neural progenitor cells, nestin, was detected using immunocytochemistry. After INPC / EGFP was induced by 50 mL / L fetal bovine serum, the cell morphology and the expression of EGFP were observed in the differentiated cells. RESULTS: The transfection efficiency of Lipofectamine 2000, TRANSfection and Sofast was (25.5 ± 2.9)%, (4.0 ± 1.7)%, (7.9 ± 1.4)% respectively, at 24 h after transfection. And Lipofectamine 2000 was the most efficient. Its transfection efficiency at 12, 24, 48 and 72 h after transfection was (17.1 ± 0.7)%, (25.5 ± 2.9)%, (19.4 ± 0.9)%, (15.6 ± 1.4)%, respectively. The expression of EGFP was the highest