Abstract
Embryonic stem cell (ESC)-derived embryoid body (EB) is a unique model for studying vascular development, in that it provides a three-dimensional microenvironment that mimics an in vivo milieu. When using gene-targeting EBs to study certain defects in vascular morphogenesis, it is necessary to determine whether the defect is due to the intrinsic loss of the gene in endothelial cells (EC) or rather due to the lack of surrounding factors that would typically promote vascular development. Here we describe a chimeric EB vessel development model, in which the utilization of the PECAM-GFP reporter gene in wild-type ESCs allows for the introduction of “normal” extracellular factors formed by its parallel differentiation to the gene-deletion EC that might otherwise be devoid of these factors.
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References
Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R (1985) The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol 87:27–45
Jakobsson L, Kreuger J, Claesson-Welsh L (2007) Building blood vessels-stem cell models in vascular biology. J Cell Biol 177:751–755
Qi Y, Liu J, Wu X, Brakebusch C, Leitges M, Han Y et al (2011) Cdc42 controls vascular network assembly through protein kinase Ciota during embryonic vasculogenesis. Arterioscler Thromb Vasc Biol 31:1861–1870
Kearney JB, Kappas NC, Ellerstrom C, DiPaola FW, Bautch VL (2004) The VEGF receptor flt-1 (VEGFR-1) is a positive modulator of vascular sprout formation and branching morphogenesis. Blood 103:4527–4535
Wu X, Li S, Chrostek-Grashoff A, Czuchra A, Meyer H, Yurchenco PD et al (2007) Cdc42 is crucial for the establishment of epithelial polarity during early mammalian development. Dev Dyn 236:2767–2778
Li S, Yurchenco PD (2006) Matrix assembly, cell polarization, and cell survival: analysis of peri-implantation development with cultured embryonic stem cells. Methods Mol Biol 329:113–125
Robson P, Stein P, Zhou B, Schultz RM, Baldwin HS (2001) Inner cell mass-specific expression of a cell adhesion molecule (PECAM-1/CD31) in the mouse blastocyst. Dev Biol 234:317–329
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Qi, Y., Saadat, S., Liu, J., Graham, A.M., Li, S. (2015). A Chimeric Embryoid Body Model to Study Vascular Morphogenesis. In: Ribatti, D. (eds) Vascular Morphogenesis. Methods in Molecular Biology, vol 1214. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1462-3_13
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DOI: https://doi.org/10.1007/978-1-4939-1462-3_13
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1461-6
Online ISBN: 978-1-4939-1462-3
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