Transfection of adherent murine peritoneal macrophages with a reporter gene using DEAE-dextran

J Immunol Methods. 1991 Nov 22;144(2):157-63. doi: 10.1016/0022-1759(91)90082-q.

Abstract

We have sought a transfection strategy which preserves cell viability and achieves sufficient efficiency to perform reporter gene assays in primary cultures of thioglycollate-elicited murine peritoneal macrophages. Murine peritoneal macrophages were transfected with an eukaryotic expression vector containing the Rous sarcoma virus enhancer-promoter upstream of the bacterial chloramphenicol acetyl-transferase reporter gene (FC4-CAT). Transfection using DEAE-dextran followed by 10% DMSO provided much higher CAT activity than either calcium-phosphate or lipofection. The transfected macrophages increased CAT activity (1.9-7.6-fold) following stimulation with 10% serum. DEAE-dextran/DMSO-mediated transfection provides a simple, inexpensive method to transfect primary cultures of adherent macrophages with heterologous plasmid DNA. Transfection of macrophages with a CAT reporter gene using this method permits the characterization of gene regulation in primary macrophage cultures in vitro.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cells, Cultured
  • Chloramphenicol O-Acetyltransferase / analysis
  • Chloramphenicol O-Acetyltransferase / genetics*
  • DEAE-Dextran / pharmacology*
  • Gene Expression Regulation, Enzymologic
  • Macrophages / enzymology*
  • Mice
  • Mice, Inbred BALB C
  • Peritoneal Cavity / cytology
  • Transfection*

Substances

  • DEAE-Dextran
  • Chloramphenicol O-Acetyltransferase