A novel method for DEAE-dextran mediated transfection of adherent primary cultured human macrophages.
J Immunol Methods
; 211(1-2): 79-86, 1998 Feb 01.
Article
em En
| MEDLINE
| ID: mdl-9617833
ABSTRACT
Primary cultured human macrophages are difficult to transfect. We have developed a DEAE-dextran DNA transfection method that mediates the reproducible transfection of primary cultured adherent human macrophages. Three factors essential for successful transfection were identified DEAE-dextran concentration, the quantity of DNA per transfection and the incubation time of the macrophages with the transfection medium. Maximum levels of luciferase expression were attained within 24 h and maintained for at least 56 h after transfection. While serum in the transfection medium attenuated transfection, the treatment of the macrophages with chloroquine, DMSO, or glycerol did not enhance transfection within this system. A CMV enhancer/promoter mediated substantially greater luciferase expression in the macrophages than either HIV or RSV LTRs. DEAE-dextran facilitated superior transfection compared to either cationic liposome and calcium phosphate methods, and was more practical compared to electroporation for multiple transfections. This transfection protocol provides a simple, inexpensive, reproducible method for the evaluation of gene expression in primary cultured adherent human macrophages.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Transfecção
/
Macrófagos
Tipo de estudo:
Guideline
/
Prognostic_studies
Limite:
Adult
/
Humans
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
1998
Tipo de documento:
Article