Skip to main content

Transplantation of Chimeric Fetal Liver to Study Hematopoiesis

  • Protocol
Hematopoietic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 430))

Summary

Complementing mutant embryos or embryonic stem cells with normal cells in embryonic chimeras is a valuable tool for investigating phenotypes. Chimera approaches provide a method to examine the phenotype of mutant cells, including hematopoiesis, in mutants with early embryonic lethality. Complementation with normal cells in a chimera can, in most instances, rescue mutant cells to later stages of gestation and beyond, permitting analysis of contribution and function of mutant cells in various organs, both within the chimera, but also by using functional transplantation assays for hematopoietic stem and progenitor cells. This chapter describes principles and methods for the generation of mouse chimeras, for identification and quantitative analysis of cell contribution in chimeras, and for chimeric fetal liver transplantation into adult recipients and analysis of mutant cells in the adult.

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 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Feil R. (2007) Conditional somatic mutagenesis in the mouse using site- specific recombinases. Handb Exp Pharmacol 178, 3–28.

    Article  CAS  PubMed  Google Scholar 

  2. Matsumoto N., Kubo A., Liu h., Akita K., Laub F., Ramirez F., Keller G., Friedman S. L. (2006) Developmental regulation of yolk sac hematopoiesis by Kruppel- like factor 6. Blood 107, 1357–1365.

    Article  CAS  PubMed  Google Scholar 

  3. Tam P. P., Rossant J. (2003) Mouse embryonic chimeras: tools for studying mammalian development. Development 130, 6155–6163.

    Article  CAS  PubMed  Google Scholar 

  4. Chen J., Lansford R., Stewart V., Young F., Alt F. W. (1993) RAG- 2- deficient blastocyst complementation: an assay of gene function in lymphocyte development. Proc Natl Acad Sci USA 90, 4528–4532.

    Article  CAS  PubMed  Google Scholar 

  5. Wang L. C., Swat W., Fujiwara Y., Davidson L., Visvader J., Kuo F., Alt F. W., Gilliland D. G., Golub T. R., Orkin S. h. (1998) The TEL/ETV6 gene is required specifically for hematopoiesis in the bone marrow. Genes Dev 12, 2392–2402.

    Article  CAS  PubMed  Google Scholar 

  6. Ernst P., Fisher J. K., Avery W., Wade S., Foy D., Korsmeyer S. J. (2004) Definitive hematopoiesis requires the mixed- lineage leukemia gene. Dev Cell 6, 437–443.

    Article  CAS  PubMed  Google Scholar 

  7. Tsang A. P., Fujiwara Y., Hom D. B., Orkin S. h. (1998) Failure of megakaryopoiesis and arrested erythropoiesis in mice lacking the GATA- 1 transcriptional cofactor FOG. Genes Dev 12, 1176–1188.

    Article  CAS  PubMed  Google Scholar 

  8. Eckardt S., Leu N. A., Bradley h. L., Kato h., Bunting K. D., Mclaughlin K. J. (2007) Hematopoietic reconstitution with androgenetic and gynogenetic stem cells. Genes Dev 21, 409–419.

    Article  CAS  PubMed  Google Scholar 

  9. Nagy A., Gertsenstein M., Vintersten K., Behringer R. (2003) Manipulating the Mouse Embryo, 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  10. Abbondanzo S. J., Gadi I., Stewart C. L. (1993) Derivation of embryonic stem cell lines, in Methods Enzymol 225: Guide to Techniques in Mouse Development (Wassarman, P.M, DePamphilis, M.L., eds.), Academic Press, San Diego, CA, pp. 803–823.

    Google Scholar 

  11. Okabe M., Ikawa M., Kominami K., Nakanishi T., Nishimune Y. (1997) ‘Green mice’ as a source of ubiquitous green cells. FEBS Lett 407, 313–319.

    Article  CAS  PubMed  Google Scholar 

  12. Kawakami N., Sakane N., Nishizawa F., Iwao M., Fukada S. I., Tsujikawa K., Kohama Y., Ikawa M., Okabe M., Yamamoto h. (1999) Green fluorescent protein- transgenic mice: immune functions and their application to studies of lymphocyte development. Immunol Lett 70, 165–171.

    Article  CAS  PubMed  Google Scholar 

  13. Spangrude G. J., Cho S., Guedelhoefer O., Vanwoerkom R. C., Fleming W. h. (2006) Mouse models of hematopoietic engraftment: limitations of transgenic green fluorescent protein strains and a high- performance liquid chromatography approach to analysis of erythroid chimerism. Stem Cells 24, 2045–2051.

    Article  CAS  PubMed  Google Scholar 

  14. Wang Z., Jaenisch R. (2004) At most three ES cells contribute to the somatic lineages of chimeric mice and of mice produced by ES- tetraploid complementation. Dev Biol 275, 192–201.

    Article  CAS  PubMed  Google Scholar 

  15. Yagi T., Tokunaga T., Furuta Y., Nada S., Yoshida M., Tsukada T., Saga Y., Takeda N., Ikawa Y., Aizawa S. (1993) A novel ES cell line, TT2, with high germline- differentiating potency. Anal Biochem 214, 70–76.

    Article  CAS  PubMed  Google Scholar 

  16. Stewart C. L. (1993) Production of chimeras between embryonic stem cells and embryos, in Methods Enzymol 225: Guide to Techniques in Mouse Development (Wassarman, P.M, DePamphilis, M.L., eds.), Academic Press, San Diego, CA, pp. 823–855.

    Google Scholar 

  17. Mann J. R. (1993) Surgical Techniques in Production of Transgenic Mice, in Methods Enzymol 225: Guide to Techniques in Mouse Development (Wassarman, P.M, DePamphilis, M.L., eds.), Academic Press, San Diego, CA, pp. 782–793.

    Google Scholar 

  18. Nagy A., Rossant J. (1993) Production of completely ES cell derived fetuses, in Gene Targeting, 1 ed. IRL (Joyner A.L., ed.), Oxford, pp. 147–178.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Eckardt, S., McLaughlin, K.J. (2008). Transplantation of Chimeric Fetal Liver to Study Hematopoiesis. In: Bunting, K.D. (eds) Hematopoietic Stem Cell Protocols. Methods in Molecular Biology™, vol 430. Humana Press. https://doi.org/10.1007/978-1-59745-182-6_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-182-6_14

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-868-3

  • Online ISBN: 978-1-59745-182-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics