Skip to main content
SpringerLink
Log in
Book cover

Stem Cell Migration pp 143–155Cite as

Whole Embryo Imaging of Hematopoietic Cell Emergence and Migration

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 750))

Abstract

The use of transgenic mice in which tissue or lineage-specific, cell-restricted promoters drive fluorescent reporters has recently been reported as a means to follow the in vivo migration of various hematopoietic cells during murine development. At present there is limited ability of these approaches to image the emergence of the first hematopoietic cell subsets due to lack of unique markers that define those hematopoietic cells. We have utilized whole embryo analysis via immunostaining and confocal laser-scanning microscopic (CLSM) imaging to define the emergence of the first hematopoietic elements in the yolk sac of the developing conceptus. The methods employed to examine yolk sac hematopoiesis may be applied to hematopoietic cell emergence in the embryo proper or fetal liver in the generation of a complete map of hematopoietic ontogeny.

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

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   159.00
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Sabin, F. R. (1920) Studies on the origin of blood-vessels and of red blood-corpuscles as seen in the living blastoderm of chicks during the second day of incubation Contr Embryol 9, 215–262.

    Google Scholar 

  2. Luckett, W. P. (1978) Origin and differentiation of the yolk sac and extraembryonic mesoderm in presomite human and rhesus monkey embryos Am J Anat 152, 59–97.

    Article  PubMed  CAS  Google Scholar 

  3. Moore, M. A., and Metcalf, D. (1970) Ontogeny of the haemopoietic system: yolk sac origin of in vivo and in vitro colony forming cells in the developing mouse embryo Br J Haematol 18, 279–96.

    Article  PubMed  CAS  Google Scholar 

  4. Rhodes, K. E., Gekas, C., Wang, Y., Lux, C. T., Francis, C. S., Chan, D. N., Conway, S., Orkin, S. H., Yoder, M. C., and Mikkola, H. K. (2008) The emergence of hematopoietic stem cells is initiated in the placental vasculature in the absence of circulation Cell Stem Cell 2, 252–63.

    Article  PubMed  CAS  Google Scholar 

  5. North, T. E., de Bruijn, M. F., Stacy, T., Talebian, L., Lind, E., Robin, C., Binder, M., Dzierzak, E., and Speck, N. A. (2002) Runx1 expression marks long-term repopulating hematopoietic stem cells in the midgestation mouse embryo Immunity 16, 661–72.

    Article  PubMed  CAS  Google Scholar 

  6. Samokhvalov, I. M., Samokhvalova, N. I., and Nishikawa, S. (2007) Cell tracing shows the contribution of the yolk sac to adult haematopoiesis Nature 446, 1056–61.

    Article  PubMed  CAS  Google Scholar 

  7. de Bruijn, M. F., Ma, X., Robin, C., Ottersbach, K., Sanchez, M. J., and Dzierzak, E. (2002) Hematopoietic stem cells localize to the endothelial cell layer in the midgestation mouse aorta Immunity 16, 673–83.

    Article  PubMed  Google Scholar 

  8. Jones, E. A., Crotty, D., Kulesa, P. M., Waters, C. W., Baron, M. H., Fraser, S. E., and Dickinson, M. E. (2002) Dynamic in vivo imaging of postimplantation mammalian embryos using whole embryo culture Genesis 34, 228–35.

    Article  PubMed  CAS  Google Scholar 

  9. Fraser, S. T., Hadjantonakis, A. K., Sahr, K. E., Willey, S., Kelly, O. G., Jones, E. A., Dickinson, M. E., and Baron, M. H. (2005) Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice Genesis 42, 162–71.

    Article  PubMed  CAS  Google Scholar 

  10. Jones, E. A., Baron, M. H., Fraser, S. E., and Dickinson, M. E. (2004) Measuring hemodynamic changes during mammalian development Am J Physiol Heart Circ Physiol 287, H1561–9.

    Article  PubMed  CAS  Google Scholar 

  11. Jones, E. A., Baron, M. H., Fraser, S. E., and Dickinson, M. E. (2005) Dynamic in vivo imaging of mammalian hematovascular ­development using whole embryo culture Methods Mol Med 105, 381–94.

    PubMed  Google Scholar 

  12. Larina, I. V., Shen, W., Kelly, O. G., Hadjantonakis, A. K., Baron, M. H., and Dickinson, M. E. (2009) A membrane associated mCherry fluorescent reporter line for studying vascular remodeling and cardiac function during murine embryonic development Anat Rec (Hoboken) 292, 333–41.

    Google Scholar 

  13. Jacobs, R. E., and Fraser, S. E. (1994) Magnetic resonance microscopy of embryonic cell lineages and movements Science 263, 681–4.

    Article  PubMed  CAS  Google Scholar 

  14. Sharpe, J., Ahlgren, U., Perry, P., Hill, B., Ross, A., Hecksher-Sorensen, J., Baldock, R., and Davidson, D. (2002) Optical projection tomography as a tool for 3D microscopy and gene expression studies Science 296, 541–5.

    Google Scholar 

  15. Walls, J. R., Sled, J. G., Sharpe, J., and Henkelman, R. M. (2007) Resolution improvement in emission optical projection tomography Phys Med Biol 52, 2775–90.

    Article  PubMed  Google Scholar 

  16. Corbel, C., and Salaun, J. (2002) AlphaIIb integrin expression during development of the murine hemopoietic system Dev Biol 243, 301–11.

    Article  PubMed  CAS  Google Scholar 

  17. Ferkowicz, M. J., Starr, M., Xie, X., Li, W., Johnson, S. A., Shelley, W. C., Morrison, P. R., and Yoder, M. C. (2003) CD41 expression defines the onset of primitive and definitive hematopoiesis in the murine embryo Development 130, 4393–403.

    Article  PubMed  CAS  Google Scholar 

  18. Hashimoto, K., Fujimoto, T., Shimoda, Y., Huang, X., Sakamoto, H., and Ogawa, M. (2007) Distinct hemogenic potential of endothelial cells and CD41+ cells in mouse embryos Dev Growth Differ 49, 287–300.

    Article  PubMed  Google Scholar 

  19. Mikkola, H. K., Fujiwara, Y., Schlaeger, T. M., Traver, D., and Orkin, S. H. (2003) Expression of CD41 marks the initiation of definitive hematopoiesis in the mouse embryo Blood 101, 508–16.

    Article  PubMed  CAS  Google Scholar 

  20. Mitjavila-Garcia, M. T., Cailleret, M., Godin, I., Nogueira, M. M., Cohen-Solal, K., Schiavon, V., Lecluse, Y., Le Pesteur, F., Lagrue, A. H., and Vainchenker, W. (2002) Expression of CD41 on hematopoietic progenitors derived from embryonic hematopoietic cells Development 129, 2003–13.

    PubMed  CAS  Google Scholar 

  21. Ody, C., Vaigot, P., Quere, P., Imhof, B. A., and Corbel, C. (1999) Glycoprotein IIb-IIIa is expressed on avian multilineage hematopoietic progenitor cells Blood 93, 2898–906.

    PubMed  CAS  Google Scholar 

  22. Tavian, M., and Peault, B. (2005) Embryonic development of the human hematopoietic system Int J Dev Biol 49, 243–50.

    Article  PubMed  CAS  Google Scholar 

  23. Palis, J., Robertson, S., Kennedy, M., Wall, C., and Keller, G. (1999) Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse Development 126, 5073–84.

    PubMed  CAS  Google Scholar 

  24. Yokomizo, T., Takahashi, S., Mochizuki, N., Kuroha, T., Ema, M., Wakamatsu, A., Shimizu, R., Ohneda, O., Osato, M., Okada, H. et al. (2007) Characterization of GATA-1(+) hemangioblastic cells in the mouse embryo Embo J 26, 184–96.

    Article  PubMed  CAS  Google Scholar 

  25. Hogan, B. L., Beddington, R. S., Costantini, F., and Lacy, E. (1994) Manipulating the mouse embryo. Plainview: Cold Spring Harbor Laboratory Press.

    Google Scholar 

Download references

Acknowledgment

Jason Byars at the Indiana Center for Biological Microscopy wrote the Image J stitching macros.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN, USA

    Mervin C. Yoder

Authors
  1. Michael J. Ferkowicz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mervin C. Yoder
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mervin C. Yoder .

Editor information

Editors and Affiliations

  1. Medical Center, Division of Experimental Hematology, Cincinnati Children's Hospital, Burnet Ave. 3333, Cincinnati, 45229, Ohio, USA

    Marie-Dominique Filippi

  2. Medical Center, Division of Experimental Hematology, Cincinnati Children's Hospital, Burnet Avenue 3333, Cincinnati, 45229, Ohio, USA

    Hartmut Geiger

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Ferkowicz, M.J., Yoder, M.C. (2011). Whole Embryo Imaging of Hematopoietic Cell Emergence and Migration. In: Filippi, MD., Geiger, H. (eds) Stem Cell Migration. Methods in Molecular Biology, vol 750. Humana Press. https://doi.org/10.1007/978-1-61779-145-1_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-145-1_10

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-144-4

  • Online ISBN: 978-1-61779-145-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation