title: Establishment of Green Fluorescent Protein and Firefly Luciferase Expressing Mouse Primary Macrophages for In Vivo Bioluminescence Imaging.
abstract: Macrophages play a key role in tissue homeostasis as well as in a range of pathological conditions including atherosclerosis, cancer, and autoimmunity. Many aspects of their in vivo behavior are, however, poorly understood. Bioluminescence imaging (BLI) with green fluorescent protein (GFP) and firefly luciferase (FLUC) labelled autologous reporter macrophages could potentially offer a powerful tool to study macrophage biology, but this approach has been hindered by the relative difficulty of efficient gene transfer into primary macrophages. Here we describe a straightforward method for producing large numbers of GFP/FLUC expressing mouse primary macrophages utilizing lentivirus vector, cyclosporine, and a double infection strategy. Using this method we achieved up to 60% of macrophages to express GFP with correspondingly high FLUC signal. When injected into the circulation using a mouse model of local biomaterial induced inflammation and osteolysis, macrophages were initially detectable within the lungs, followed by systemic homing to the local area of chronic inflammation in the distal femur. In addition, transduced macrophages maintained their ability to assume M1 and M2 phenotypes although the GFP/FLUC expression was altered by the polarizing signals. These reporter macrophages could prove to be valuable tools to study the role of macrophages in health and disease.
authors.affiliation: Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Jukka
authors.initials: J
authors.lastname: Pajarinen
authors.affiliation: Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Tzu-Hua
authors.initials: TH
authors.lastname: Lin
authors.affiliation: Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Taishi
authors.initials: T
authors.lastname: Sato
authors.affiliation: Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Florence
authors.initials: F
authors.lastname: Loi
authors.affiliation: Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Yao
authors.affiliation: Department of Medicine, Institute of Clinical Medicine, University of Helsinki, Helsinki, Finland.
authors.forename: Yrjö T
authors.initials: YT
authors.lastname: Konttinen
authors.affiliation: Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Stuart B
authors.initials: SB
authors.lastname: Goodman
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D003911:Dextrans / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006583:Hexadimethrine Bromide / Q000494:pharmacology; D049409:Luciferases, Firefly / Q000235:genetics*; D049449:Luminescence; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D057054:Molecular Imaging; D014161:Transduction, Genetic
chemical_list: D003911:Dextrans; D049452:Green Fluorescent Proteins; D006583:Hexadimethrine Bromide; D016572:Cyclosporine; D049409:Luciferases, Firefly