High-Efficiency Gene Transfection of Macrophages by Lipoplexes

Details

• Personal Author:
  Dokka S ; Rojanasakul Y ; Shi XL ; Toledo D ; Ye JP
• Description:
  Macrophage transfection studies are crucial for understanding gene regulation and expression. However, gene transfection in macrophages is difficult. We have shown here that macrophages are more resistant to gene transfection compared with other cell types. To further develop an efficient gene delivery system for macrophages, we evaluated various liposomal and non-liposomal agents including LipofectAMINE(R), Lipofectin(R), DOTAP, DEAE-dextran, and the DNA condensing agent protamine sulfate for their ability to promote gene transfection. CMV-luciferase was used as a reporter plasmid. Macrophage transfection was maximal at the DNA:LipofectAMINE:protamine ratio of 1:12:1 mu g/ml The LipofectAMINE formulation showed a 10-12-fold increase in transfection efficiency over DOTAP and a 4-5-fold increase over Lipofectin. This transfection method showed minimal toxicity at the concentrations tested and was at least 20-25-fold superior to the most frequently used DEAE-dextran method for macrophage transfection. [Description provided by NIOSH]
• Subjects:
  Genetics Occupational Health Safety
• Keywords:
  Analytical-methods Analytical-processes Cell-biology Cell-metabolism Cell-transformation Genes Genetic-factors Genotoxic-effects
• ISSN:
  0378-5173
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  Oregon ; OSHA Region 10 ; OSHA Region 3 ; OSHA Region 5 ; West Virginia ; Wisconsin
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  97-104
• Volume:
  206
• Issue:
  1
• NIOSHTIC Number:
  nn:20032339
• Citation:
  Int J Pharm 2000 Sep; 206(1-2):97-104
• Email:
  yrojanasakul@hsc.wvu.edu
• Federal Fiscal Year:
  2000
• Peer Reviewed:
  True
• Source Full Name:
  International Journal of Pharmaceutics
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:a01ad2bf07026fb243fccee3e51c12f0e074fd7d51fb5a1ed3e9272f435f4e0796e4dafd6d0f90a7697602bc3a81818334de0d947bcb4945c3a9c3e5a659bfcc
• Download URL:
  https://stacks.cdc.gov/view/cdc/186357/cdc_186357_DS1.pdf
• File Type:
  [PDF - 84.65 KB ]