Transfection of human peripheral blood mononuclear cells using immunoporation.
Immunoporation has been found to be able to efficiently transfect a wide range of human cultured cell lines. This report shows that peripheral blood mononuclear cells can also be efficiently transfected using immunoporation. The immunoporation of the cells with fluorescent TMR-Dextran, using Immunofect MG beads, indicates that transient holes of 5.4nm in diameter or larger are formed during immunoporation. The efficiencies of transfection of lymphocytes transfected with vectors coding for EGFP and lacZ were found to be within the range of 15-30% with high levels of cell viability of more than 90%. In addition, it was observed that mononuclear cells stimulated with PHA expressed transfected reporter genes with a higher efficiency. In conclusion, these results demonstrate that immunoporation using Immunofect MG beads can be used for the efficient transfection of primary lymphocytes with DNA or other macromolecules.
Nateri
A S
AS
Department of Biological Sciences, University of Essex, Colchester CO4 3SQ, UK.
Tzavelas
C
C
Bildirici
L
L
Rickwood
D
D
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D002448:Cell Adhesion / Q000276:immunology; D002463:Cell Membrane Permeability; D003911:Dextrans / Q000737:chemistry / Q000378:metabolism; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics / Q000493:pharmacokinetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008863:Microspheres; D010835:Phytohemagglutinins / Q000494:pharmacology; D012235:Rhodamines / Q000737:chemistry; D013314:Stress, Mechanical; D014162:Transfection / Q000295:instrumentation / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D000906:Antibodies; D003911:Dextrans; D010835:Phytohemagglutinins; D012235:Rhodamines; C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins; C005358:tetramethylrhodamine; D001616:beta-Galactosidase