Transfection of recombinant human NT-3 gene into mouse embryonic stem cell and its expression in vitro / 中华神经医学杂志

ABSTRACT

Objective To investigate the possibility of in vitro expression of human neurotrophin-3 (NT-3) gene in mouse embryonic stem (ES) cells. Methods The reconlbinant eukaryotic expression vector,PCDNA-3.1(+)-NT-3 was transiently transfected into ES cells by means of liposome mediated method. NT-3 protein and mRNA were tested by immunohistochemical method and RT-PCR,respectively.NT-3 protein secretion in the supernatant of cell culture was detected by ELISA.Results By immunohistochemistry, ES cells were red stained in cytoplasma, showing that the transfected NT-3 plasmid was expressed in the ES cells.RT-PCR got a 200 bp segment in the transfected cells,and the NT-3 protein secretion in the supernatrdnt of transfected cell culture detected by ELISA was dramatically higher than that of control group(P＜0.05). Conclusions Successful transfection of recombinant PCDNA-3.1(+)-NT-3 into the ES cells that leads to stable expression of NT-3 gene can be a potential effective gene delivery approach in the treatnlent of Parkinson's disease.