[Transfection of human 293 and CD34+ cells with recombinant green fluorescent protein adeno- associated virus].
OBJECTIVE
To investigate the expressions of recombinant green fluorescent protein (GFP) in 293 cells (human embryonic kidney cells) and CD34(+) cells transfected with adeno-associated virus carrying the gene encoding the recombinant GFP.


METHODS
Mononuclear cells from the bone marrow were collected on cell separator CS-3000, and CD34(+) cells were separated on immunomagnetic MiniMACS columns. CD34(+) cells and 293 cells were transfected with recombined adeno-associated virus respectively, and the expressions of the GFP were detected by flow cytometry and fluorescence microscopy.


RESULTS
GFP expression was observed in the two cells under fluorescent microscope. The highest transfection efficiency of the recombined adeno-associated virus was about 32.8% in 293 cells and 25% in CD34(+) cells, and decreased with the prolongation of transfection time.


CONCLUSIONS
Adeno-associated viral vector encoding recombinant GFP can be transfected into human 293 cells and CD34(+) cells, which provides a basis for future gene therapy.
Li
Li-bo
LB
Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Feng
Ru
R
Zhou
Shu-yun
SY
D018952:Antigens, CD34 / Q000378:metabolism*; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004622:Embryo, Mammalian; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000166:cytology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D018952:Antigens, CD34; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins