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Generation of Innate Immune Reporter Cells Using Retroviral Transduction

  • Yamel Cardona Gloria
  • Eicke Latz
  • Dominic De NardoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1714)

Abstract

Innate immune cells are notoriously difficult to transfect; however, retroviruses can be used to stably integrate genes of interest into the host genome of primary or immortalized immune cells resulting in the generation of reporter cells. Here, we provide a detailed protocol covering the production of retroviruses, retroviral infection of innate immune target cells (including isolation and differentiation of murine bone marrow cells to macrophages), and several methods for enrichment of positively transduced cells.

Keywords

Retrovirus Infection Transduction Innate immune reporter cells Bone marrow isolation Bone marrow-derived macrophages Fluorescent protein Innate immunity 

Notes

Acknowledgements

We thank R. Stahl (Institute of Innate Immunity, Bonn) for assistance in generating the murine IRAK4-mCitrine and control mCitrine retroviral plasmids. This work was supported in part by the intramural BONFOR research support at the University of Bonn (D.D).

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Yamel Cardona Gloria
    • 1
  • Eicke Latz
    • 2
    • 3
    • 4
  • Dominic De Nardo
    • 5
    • 6
    Email author
  1. 1.Department of Immunology, Interfaculty Institute for Cell BiologyUniversity of TübingenTübingenGermany
  2. 2.Institute of Innate ImmunityUniversity Hospital, University of BonnBonnGermany
  3. 3.Department of Infectious Diseases and ImmunologyUniversity of Massachusetts Medical SchoolWorcesterUSA
  4. 4.German Center for Neurodegenerative DiseasesBonnGermany
  5. 5.Inflammation DivisionThe Walter & Eliza Hall Institute of Medical ResearchParkvilleAustralia
  6. 6.Department of Medical BiologyThe University of MelbourneParkvilleAustralia

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