Abstract
Embryonic stem (ES) cells are pluripotent cells derived from blastocyst-stage embryos. They are characterized by their infinite self-renewal capacity and their ability to differentiate into many cell types in vitro as well as in vivo. The present protocol describes culture conditions that allow efficient differentiation of mouse ES cells toward the endothelial lineage, both in two-dimensional cultures and in three-dimensional, multicellular embryoid bodies. We also provide a protocol for establishing recombinant ES cell clones, giving the example of cells expressing green fluorescent protein and an antibiotic resistance gene under the control of an endothelial-specific promoter. These transgenes allow the visualization and the genetic selection of endothelial cells from a heterogeneous population of differentiating ES cells. Potential applications for these differentiation models are given, including the study of the endothelial cell lineage and its derivatives, the testing of molecules involved in angiogenesis, and the potential use of selected endothelial cells in vivo.
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
Mummery C. L., Feijen A., Freund E., and Shen S. (1990) Characteristics of embryonic stem cell differentiation: a comparison with two embryonal carcinoma cell lines. Cell Differ. Dev. 30, 1–18.
Marchetti S., Gimond C., Iljin K., et al. (2002) Endothelial cells genetically selected from differentiating mouse embryonic stem cells incorporate at sites of neovascularization in vivo. J. Cell Sci. 115, 2075–2085.
Vittet D., Prandini M. H., Berthier R., et al. (1996) Embryonic stem cells differentiate in vitro to endothelial cells through successive maturation steps. Blood 88, 3424–3431.
Risau W., Sariola H., Zerwes. G., et al. (1988) Vasculogenesis and angiogenesis in embryonic-stem-cell-derived embryoid bodies. Development 102, 471–478.
Hirashima M., Kataoka H., Nishikawa S., and Matsuyoshi N. (1999) Maturation of embryonic stem cells into endothelial cells in an in vitro model of vasculogenesis. Blood 93, 1253–1263.
Levenberg S., Golub J. S., Amit M., Itskovitz-Eldor J., and Langer R. (2002) Endothelial cells derived from human embryonic stem cells. Proc. Natl. Acad. Sci. USA 99, 4391–4396.
Magnusson P., Rolny C., Jakobsson L., et al. (2004) Deregulation of Flk-1/vascular endothelial growth factor receptor-2 in fibroblast growth factor receptor-1-deficient vascular stem cell development. J. Cell Sci. 117, 1513–1523.
Sone M., Itoh H., Yamashita J., et al. (2003) Different differentiation kinetics of vascular progenitor cells in primate and mouse embryonic stem cells. Circulation 107, 2085–2088.
Gasmi A., Bourcier C., Aloui Z., et al. (2002) Complete structure of an increasing capillary permeability protein (ICPP) purified from Vipera lebetina venom. ICPP is angiogenic via vascular endothelial growth factor receptor signaling. J. Biol. Chem. 277,29,992–29,998.
Deroanne C. F., Bonjean K., Servotte S., et al. (2002) Histone deacetylases inhibitors as antiangiogenic agents altering vascular endothelial growth factor signaling. Oncogene 21, 427–436.
Yamashita J., Itoh H., Hirashima M., et al. (2000) Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors. Nature 408, 92–96.
Ishida A., Murray J., Saito Y., et al. (2001) Expression of vascular endothelial growth factor receptors in smooth muscle cells. J. Cell Physiol. 188, 359–368.
Nakayama N., Duryea D., Manoukian R., Chow G., and Han C. Y. (2003) Macroscopic cartilage formation with embryonic stem-cell-derived mesodermal progenitor cells. J. Cell Sci. 116, 2015–2028.
Zhang H., Vutskits L., Pepper M. S., and Kiss J. Z. (2003) VEGF is a chemoattractant for FGF-2-stimulated neural progenitors. J. Cell Biol. 163}, 1375–1384.
Korhonen J., Lahtinen I., Halmekyto M., et al. (1995) Endothelial-specific gene expression directed by the tie gene promoter in vivo. Blood 86, 1828–1835.
Iljin K., Dube A., Kontusaari S., et al. (1999) Role of ets factors in the activity and endothelial cell specificity of the mouse Tie gene promoter. FASEB J. 13, 377–386.
Arciniegas E., Sutton A. B., Allen T. D., and Schor A. M. (1992) Transforming growth factor beta 1 promotes the differentiation of endothelial cells into smooth muscle-like cells in vitro. J. Cell Sci. 103, 521–529.
Hayashi K., Saga H., Chimori Y., Kimura K., Yamanaka Y., and Sobue K. (1998) Differentiated phenotype of smooth muscle cells depends on signaling pathways through insulin-like growth factors and phosphatidylinositol 3-kinase. J. Biol. Chem. 273, 28,860–28,867.
Ema M., Faloon P., Zhang W. J., et al. (2003) Combinatorial effects of Flk1 and Tal1 on vascular and hematopoietic development in the mouse. Genes Dev. 17, 380–393.
Watabe T., Nishihara A., Mishima K., et al. (2003) TGF-beta receptor kinase inhibitor enhances growth and integrity of embryonic stem cell-derived endothelial cells. J. Cell Biol. 163, 1303–1311.
Yurugi-Kobayashi T., Itoh H., Yamashita J., et al. (2003) Effective contribution of transplanted vascular progenitor cells derived from embryonic stem cells to adult neovascularization in proper differentiation stage. Blood 101, 2675–2678.
Shen G., Tsung H. C., Wu C. F., et al. (2003) Tissue engineering of blood vessels with endothelial cells differentiated from mouse embryonic stem cells. Cell Res. 13, 335–341.
Brunet A., Pagés G., and Pouysségur J. (1994) Constitutively active mutants of MAP kinase kinase (MEK1) induce growth factor-relaxation and oncogenicity when expressed in fibroblasts. Oncogene 9, 3379–3387.
Burdon T., Stracey C., Chambers I., Nichols J., and Smith A. (1999) Suppression of SHP-2 and ERK signalling promotes self-renewal of mouse embryonic stem cells. Dev. Biol. 210, 30–43.
Kosuga M., Enosawa S., Li S. K., et al. (2000) Strong, long-term transgene expression in rat liver using chicken beta-actin promoter associated with cytomegalovirus immediateearly enhancer (CAG promoter). Cell Transplant. 9, 675–680.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Gimond, C., Marchetti, S., Pagés, G. (2006). Differentiation of Mouse Embryonic Stem Cells Into Endothelial Cells. In: Turksen, K. (eds) Embryonic Stem Cell Protocols. Methods in Molecular Biology™, vol 330. Humana Press. https://doi.org/10.1385/1-59745-036-7:303
Download citation
DOI: https://doi.org/10.1385/1-59745-036-7:303
Publisher Name: Humana Press
Print ISBN: 978-1-58829-784-6
Online ISBN: 978-1-59745-036-2
eBook Packages: Springer Protocols