title: In vitro and in vivo transfection of primary phagocytes via microbubble-mediated intraphagosomal sonoporation.
abstract: The professional phagocytes, such as macrophages and dendritic cells, are the subject of numerous research efforts in immunology and cell biology. The use of primary phagocytes in these investigations however, are limited by their inherent resistance to transfection with DNA constructs. As a result, the use of phagocyte-like immortalized cell lines is widespread. While these cell lines are transfection permissive, they are generally regarded as poor biological substitutes for primary phagocytes. By exploiting the phagocytic machinery of primary phagocytes, we developed a non-viral method of DNA transfection of macrophages that employs intraphagosomal sonoporation mediated by internalized lipid-based microbubbles. This approach enables the transfection of primary phagocytes in vitro, with a modest, but reliable efficiency. Furthermore, this methodology was readily adapted to transfect murine peritoneal macrophages in vivo. This technology has immediate application to current research efforts and has potential for use in gene therapy and vaccination strategies.
authors.affiliation: Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, AB, Canada.
authors.forename: Jason C M
authors.initials: JC
authors.lastname: Lemmon
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: McFarland
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Rybicka
authors.forename: Dale R
authors.initials: DR
authors.lastname: Balce
authors.forename: Kyle R
authors.initials: KR
authors.lastname: McKeown
authors.forename: Regina M
authors.initials: RM
authors.lastname: Krohn
authors.forename: Terry O
authors.initials: TO
authors.lastname: Matsunaga
authors.forename: Robin M
authors.initials: RM
authors.lastname: Yates
mesh_terms: D000818:Animals; D002460:Cell Line; D005434:Flow Cytometry; D066298:In Vitro Techniques; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D045423:Microbubbles; D010586:Phagocytes / Q000166:cytology / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics