Skip to main content

Generation and Characterization of MacGreen Mice, the Cfs1r-EGFP Transgenic Mice

  • Protocol
  • First Online:
Leucocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 844))

Abstract

Macrophage colony-stimulating factor (CSF-1) regulates the differentiation, proliferation, and survival of cells of the mononuclear phagocyte system. The activity of CSF-1 is mediated by the CSF-1 receptor (CSFlR, CD115) that is encoded by c-fms (Csf1r) protooncogene. The c-fms gene is expressed in macrophage, trophoblast cell lineages, and to some extent granulocytes. A reporter gene construct containing 3.5-kb of 5′ flanking sequence and the downstream intron 2 of the c-fms gene directed expression of enhanced green fluorescent protein (EGFP) to cells expressing the c-fms gene including the macrophages and trophoblasts. EGFP was detected in trophoblasts from the earliest stage of implantation. During embryonic development, EGFP expression highlighted the large numbers of c-fms positive macrophages in most organs. These embryonic macrophages contribute to organogenesis and tissue remodeling. In adult c-fms EGFP transgenic mice, which have been called the MacGreen mice, EGFP expressed in all tissue macrophage populations and permitted convenient detection of tissue macrophages as well as facilitates their isolation from various tissues.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Sasmono, R. T., Oceandy, D., Pollard, J. W. et al. (2003) A macrophage colony-stimulating factor receptor-green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse, Blood 101, 1155–1163.

    Article  PubMed  CAS  Google Scholar 

  2. Sasmono, R. T., Ehrnsperger, A., Cronau, S. L. et al. (2007) Mouse neutrophilic granulocytes express mRNA encoding the macrophage colony-stimulating factor receptor (CSF-1R) as well as many other macrophage-specific transcripts and can transdifferentiate into macrophages in vitro in response to CSF-1, J Leukoc Biol 82, 111–123.

    Article  PubMed  CAS  Google Scholar 

  3. Yue, X., Favot, P., Dunn, T. L. et al. (1993) Expression of mRNA encoding the macrophage colony-stimulating factor receptor (c-fms) is controlled by a constitutive promoter and tissue-specific transcription elongation, Mol Cell Biol 13, 3191–3201.

    PubMed  CAS  Google Scholar 

  4. Gordon, J. W., Scangos, G. A., Plotkin, D. J. et al. (1980) Genetic transformation of mouse embryos by microinjection of purified DNA, Proc Natl Acad Sci USA 77, 7380–7384.

    Article  PubMed  CAS  Google Scholar 

  5. Awasthi, A., and Kuchroo, V. K. (2009) Th17 cells: from precursors to players in inflammation and infection, Int Immunol 21, 489–498.

    Article  PubMed  CAS  Google Scholar 

  6. Nagy, A., Gertsenstein, M., Vintersten, K.et al. (2003) Manipulating the Mouse Embryo: A Laboratory Manual 3ed., Cold Spring Harbor Laboratory Press.

    Google Scholar 

  7. Hunter, C. A., Roberts, C. W., Alexander, J. (1992) Kinetics of cytokine mRNA production in the brains of mice with progressive toxoplasmic encephalitis, European Journal of Immunology 22, 2317–2322.

    Article  PubMed  CAS  Google Scholar 

  8. Sherr, C. J. (1990) Colony-stimulating factor-1 receptor, Blood 75, 1–12.

    PubMed  CAS  Google Scholar 

  9. Albieri, A., Bevilacqua, E. (1996) Induction of erythrophagocytic activity in cultured mouse trophoblast cells by phorbol myristate acetate and all-trans-retinal, Placenta 17, 507–512.

    Article  PubMed  CAS  Google Scholar 

  10. Hume, D. A., Gordon, S. (1983) Optimal conditions for proliferation of bone marrow-derived mouse macrophages in culture: the roles of CSF-1, serum, Ca2+, and adherence, J Cell Physiol 117, 189–194.

    Article  PubMed  CAS  Google Scholar 

  11. Tushinski, R. J., Oliver, I. T., Guilbert, L. J. et al. (1982) Survival of mononuclear phagocytes depends on a lineage-specific growth factor that the differentiated cells selectively destroy, Cell 28, 71–81.

    Article  PubMed  CAS  Google Scholar 

  12. Holt, P. G. (1979) Alveolar macrophages. I. A simple technique for the preparation of high numbers of viable alveolar macrophages from small laboratory animals, J Immunol Methods 27, 189–198.

    Google Scholar 

  13. Pavli, P., Woodhams, C. E., Doe, W. F. et al. (1990) Isolation and characterization of ­antigen-presenting dendritic cells from the mouse intestinal lamina propria, Immunology 70, 40–47.

    PubMed  CAS  Google Scholar 

  14. Sasmono, R. T., Hume, D. A. (2004) The Biology of Macrophages, in The Innate Immunity Response to Infection (Kaufmann, S. E., Medzhitov, R., and Gordon, S., Eds.), The American Society of Microbiology press.

    Google Scholar 

  15. Price, A. A., Cumberbatch, M., Kimber, I. et al. (1997) Alpha 6 integrins are required for Langerhans cell migration from the epidermis, J Exp Med 186, 1725–1735.

    Article  PubMed  CAS  Google Scholar 

  16. Sambrook, J., and Russel, D. W. (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press,, Cold Spring Harbour, New York.

    Google Scholar 

  17. Zeller, R. (2001) Fixation, embedding, and sectioning of tissues, embryos, and single cells, Curr Protoc Pharmacol doi: 10.1002/0471141755.pha03ds07.

Download references

Acknowledgments

We would like to thank Professor David A. Hume for his supervision during the course of this study and the Institute for Molecular Bioscience, the University of Queensland, Brisbane, Australia for providing facilities for the generation and characterization of the MacGreen mice.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R. Tedjo Sasmono .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Sasmono, R.T., Williams, E. (2012). Generation and Characterization of MacGreen Mice, the Cfs1r-EGFP Transgenic Mice. In: Ashman, R. (eds) Leucocytes. Methods in Molecular Biology, vol 844. Humana Press. https://doi.org/10.1007/978-1-61779-527-5_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-527-5_11

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-526-8

  • Online ISBN: 978-1-61779-527-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics