Abstract
The yolk sac is the initial site of hematopoiesis in the mammalian embryo. As the embryo develops, blood vessels form around primitive erythroblasts to connect the yolk sac to the embryo, delivering newly formed blood cells to the embryonic circulation. The limited accessibility of the mammalian embryo has made it difficult to study the dynamic changes in cellular development during the formation of the early hematovascular system. Therefore, we have developed a culture system for studying early hematopiesis, vasculogenesis. and angiogenesis in the mouse embryo. Early embryos (E7.5–E9.5) can be grown on the microscope stage to study the dynamics as vessels form and circulation begins. In addition, this mouse embryo culture system provides an excellent model for understanding the interplay between flow dynamics and cellular development.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Baron, M. (2001) Induction of embryonic hematopoietic and endothelial stem/progenitor cells by hedgehog-mediated signals. Differentiation 68, 175–185.
Palis, J. and Yoder, M. C. (2001) Yolk-sac hematopoiesis: the first blood cells of mouse and man. Exp. Hematol. 29, 927–936.
Yancopoulos, G. D., Klagsbrun, M., and Folkman, J. (1998) Vasculogenesis, angiogenesis, and growth factors: ephrins enter the fray at the border. Cell 93, 661–664.
Haar, J. L. and Ackerman, G. A. (1971). A phase and electron microscopic study of vasculogenesis and erythropoiesis in the yolk sac of the mouse. Anat. Rec. 170, 199–223.
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, 228–235.
Hogan, B., et al. (1994) Manipulating the Mouse Embryo: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, p. 497.
Nagy, A. (2003) Manipulating the Mouse Embryo: A Laboratory Manual, 3rd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, p. 764.
Kulesa, P. S. and Fraser, S. E. (1998) Neural crest cell dynamics revealed by time-lapse video microscopy of whole embryo chick explant cultures. Dev. Biol. 204, 327–344.
Copp, A. J. and Cockroft, D. L. (1990) Postimplantation Mammalian Embryos: A Practical Approach. The Practical approach series. IRL Press, Oxford, p. 357.
Dyer, M. A., et al. (2001) Indian hedgehog activates hematopoiesis and vasculogenesis and can respecify prospective neurectodermal cell fate in the mouse embryo. Development 128, 1717–1730.
Hadjantonakis, A. K., Dickinson, H. E., Fraser, S. E., and Papaioannov, V. E. (2003) Technicolour Transgenics: Imaging Tools for Functional Genomics in the Mouse. Nat. Rev. Genet. 4, 613–625.
Motoike, T., Loughna, S., Perens, E., Roman, B. L., Liao, W., Chau, T. C., et al. (2000) Universal GFP reporter for the study of vascular development. Genesis 28, 75–81.
Iljin, K., Petrova, T. V., Veikkola, T. V., Kumar, V., Poutanen, M., and Altitalo, K. (2002) A fluorescent Tie1 reporter allows monitoring of vascular development and endothelial cell isolation from transgenic mouse embryos. FASEB J. 16, 1764–1774.
Minegishi, N., Ohta, J., Yamagiwa, H., Suzuki, N., Kawauchi, S., Zhou, Y., et al. (1999) The mouse GATA-2 gene is expressed in the para-aortic splanchnopleura and aorta-gonads and mesonephros region. Blood 93, 4196–4207.
Hirai, H., Ogawa, M., Suzuki, N., Yamamoto, M., Breler, G., Mazda, O., et al. (2003) Hemogenic and nonhemogenic endothelium can be distinguished by the activity of fetal liver kinase (Flk)-1 promoter/enhancer during mouse embryogenesis. Blood 101, 886–893.
Bkaily, G., Pothier, P., D’Orleans-Juste, P., Simaan, M., Jacques, D., Jaalouk, D., et al. (1997) The use of confocal microscopy in the investigation of cell structure and function in the heart, vascular endothelium and smooth muscle cells. Mol. Cell. Biochem. 172, 171–194.
Quinlan, G. A., Trainor, P. A., and Tam, P. P. (1999) Cell grafting and labeling in postimplantation mouse embryos. Methods Mol. Biol. 97, 41–59.
Fahraeus, R. and Lindqvist, T. (1931) The viscosity of blood in narrow capillary tubes. Am. J. Physiol. 99, 563–568.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Jones, E.A.V., Baron, M.H., Fraser, S.E., Dickinson, M.E. (2005). Dynamic In Vivo Imaging of Mammalian Hematovascular Development Using Whole Embryo Culture. In: Baron, M.H. (eds) Developmental Hematopoiesis. Methods in Molecular Medicine, vol 105. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-826-9:381
Download citation
DOI: https://doi.org/10.1385/1-59259-826-9:381
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-296-4
Online ISBN: 978-1-59259-826-7
eBook Packages: Springer Protocols