Abstract
Interaction between endothelial cells and mural cells (pericytes and vascular smooth muscle) is essential for vascular development and maintenance1,2,3,4. Endothelial cells arise from Flk1-expressing (Flk1+) mesoderm cells5, whereas mural cells are believed to derive from mesoderm, neural crest or epicardial cells and migrate to form the vessel wall6,7,8. Difficulty in preparing pure populations of these lineages has hampered dissection of the mechanisms underlying vascular formation. Here we show that Flk1+ cells derived from embryonic stem cells can differentiate into both endothelial and mural cells and can reproduce the vascular organization process. Vascular endothelial growth factor promotes endothelial cell differentiation, whereas mural cells are induced by platelet-derived growth factor-BB. Vascular cells derived from Flk1+ cells can organize into vessel-like structures consisting of endothelial tubes supported by mural cells in three-dimensional culture. Injection of Flk1+ cells into chick embryos showed that they can incorporate as endothelial and mural cells and contribute to the developing vasculature in vivo. Our findings indicate that Flk1+ cells can act as ‘vascular progenitor cells’ to form mature vessels and thus offer potential for tissue engineering of the vascular system.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Suri, C. et al. Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell 87, 1171–1180 (1996).
Folkman, J. & D'Amore, P. A. Blood vessel formation: What is its molecular basis? Cell 87, 1153– 1155 (1996).
Darland, D. C. & D'Amore, P. A. Blood vessel maturation: vascular development comes of age. J. Clin. Invest. 103, 157–158 (1999).
Carmeliet, P. Mechanisms of angiogenesis and arteriogenesis. Nature Med. 6, 389–395 (2000).
Yamaguchi, T. P., Dumont, D. J., Conlon, R. A., Breitman, M. L. & Rossant, J. Flk1, a flt-related receptor tyrosine kinase is an early marker for endothelial precursors. Development 118, 489–498 ( 1993).
Topouzis, S. & Majesky, M. W. Smooth muscle lineage diversity in the chick embryo–Two types of aortic smooth muscle cell differ in growth and receptor-mediated transcriptional responses to transforming growth factor-β. Dev. Biol. 178, 430– 445 (1996).
Jiang, X., Rowitch, D. H., Soriano, P., McMahon, A. P. & Sucov, H. M. Fate of the mammalian cardiac neural crest. Development 127, 1607– 1616 (2000).
Mikawa, T. & Gourdie, R. G. Pericardial mesoderm generates a population of coronary smooth muscle cells migrating into the heart along with in growth of the epicardial organ. Dev. Biol. 174, 221–232 (1996).
Shalaby, F., Rosant, J., Yamaguchi, T. P., Gertsenstein, M. & Wu, X. F. Failure of blood-island formation and vasculogenesis in Flk1-deficient mice. Nature 376 , 62–66 (1995).
Eichmann, A. et al. Ligand-dependent of the endothelial and hematopoietic lineages from embryonic mesodermal cells expressing vascular endothelial growth factor 2. Proc. Natl Acad. Sci. USA 94, 5141– 5146 (1997).
Hirashima, M., Kataoka, H., Nishikawa, S., Matsuyoshi, N. & Nishikawa, S. I. Maturation of embryonic stem cells into endothelial cells in an in vitro model of vasculogenesis. Blood 93, 1253–1263 (1999).
Nishikawa, S. I., Nishikawa, S., Hirashima, M., Matsuyoshi, N. & Kodama, H. Progressive lineage analysis by cell sorting and culture identifies FLK1+VE-cadherin+ cells at a diverging point of endothelial cells and hemopoietic lineages. Development 125, 1747– 1757 (1998).
Ogawa, M. et al. Expression of α4-integrin defines the earliest precursor of hematopoietic cell lineage diverged from endothelial cells. Blood 93, 1168–1177 ( 1999).
Ueda, M., Becker, A. E., Naruko, T. & Kojima, A. Smooth muscle cell de-differentiation is a fundamental change preceding wound healing after percutaneous transluminal coronary angioplasty in humans. Coron. Artery Dis. 6, 71–81 (1995).
Hellström, M., Kalin, M., Lindahl, P., Abramsson, A. & Betsholtz, C. Role of PDGF-B and PDGFR-β in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. Development 126, 3047– 3055 (1999).
Soker, S., Takashima, S., Miao, H. Q., Neufeld, G. & Klaqsbrun, M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92, 735–745 (1998).
Lindahl, P., Johansson, B., Levéen, P. & Betsholtz, C. Pericyte loss and microaneurysm formation in PDGF-B-deficient mice. Science 277, 242–245 ( 1997).
Hirschi, K. K., Rohovsky, S. A., Beck, L. H., Smith, S. R. & D'Amore, P. A. Endothelial cells modulate the proliferation of mural cells precursors via platelet-derived growth factor-BB and heterotypic cell contact. Circ. Res. 84, 298–305 (1999).
Benjamin, L. E., Hemo, I. & Keshet, E. A plasticity window for blood vessel remodelling is defined by pericyte coverage of the preformed endothelial network and is regulated by PDGF-B and VEGF. Development 125, 1591 –1598 (1998).
DeRuiter, M. C. et al. Embryonic endothelial cells transdifferentiate into mesenchymal cells expressing smooth muscle actins in vivo and in vitro. Circ. Res. 80, 444–451 ( 1997).
Hirschi, K. K., Rohovsky, S. A. & D'Amore, P. A. PDGF, TGF-beta, and heterotypic cell-cell interactions mediate endothelial cell-induced recruitment of 10T1/2 cells and their differentiation to a smooth muscle fate. J. Cell. Biol. 141, 805–814 (1998).
Cuevas, P. et al. Pericyte endothelial gap junctions in human cerebral capillaries. Anat. Embryol. 170, 155– 159 (1984).
Choi, K., Kennedy, M., Kazarov, A., Papadimitriou, J. C. & Keller, G. A common precursor for hematopoietic and endothelial cells. Development 125, 725– 732 (1998).
Takakura, N., Kodama, H., Nishikawa, S. & Nishikawa, S. I. Preferential proliferation of murine colony-forming units in culture in a chemically defined condition with a macrophage colony-stimulating factor-negative stromal cell clone. J. Exp. Med. 184, 2301 –2309 (1996).
Shirayoshi, Y., Nose, A., Iwasaki, K. & Takeichi, M. N-linked oligosaccharides are not involved in the function of a cell-cell binding glycoprotein E-cadherin. Cell Struct. Funct. 11, 245– 252 (1986).
Kataoka, H. et al. Expressions of PDGF-receptor alpha, c-kit, and FLK1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells. Dev. Growth Differ. 39, 729– 740 (1997).
Matsuyoshi, N. et al. In vivo evidence of the critical role of cadherin-5 in murine vascular integrity. Proc. Assoc. Am. Physicians 109, 362–371 (1997).
Yoshida, H. et al. IL-7 receptor α+ CD3- cells in the embryonic intestine induces the organizing center of Peyer's patches. Int. Immunol. 11, 643– 655 (1999).
Isobe, S., Chen, S. T., Nakane, P. K. & Brown, W. R. Studies on translocation of immunoglobulins across intestinal epithelium. I. Improvements to study the peroxidase-labeled antibody method for application to study of human intestinal mucosa. Acta Histochem. Cytochem. 10, 161–171 ( 1977).
Yamamoto, T. et al. Repopulation of murine Kupffer cells after intravenous administration of liposome-encapsulated dichloromethylene diphosphonete. Am. J. Pathol. 149, 1271–1286 ( 1996).
Acknowledgements
We thank M. J. Evans for CCE ES cells, N. Matsuyoshi for the hybridoma, A. Nagafuchi for antibodies, T. Kunisada for LacZ construct, R. Yu for critical reading of the manuscript, and many of our colleagues for suggestions and discussion. This work was supported by grants from the Ministry of Education, Science, Sports and Culture of Japan, the Ministry of Health and Welfare of Japan (S.I.N.), Japanese Society for the Promotion of Science “Research for the Future” Program (H.I., M.O. and S.I.N.), Japan Tobacco Foundation, Japan Hearth Foundation & Pfizer Pharmaceuticals Grant for Research on Coronary Artery Disease and Tanabe Medical Frontier Conference. J.Y. is a recipient of the Research Fellowship grant of the Japan Society for the Promotion of Science for Young Scientists.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yamashita, J., Itoh, H., Hirashima, M. et al. Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors. Nature 408, 92–96 (2000). https://doi.org/10.1038/35040568
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/35040568
This article is cited by
-
Generation of multilineage liver organoids with luminal vasculature and bile ducts from human pluripotent stem cells via modulation of Notch signaling
Stem Cell Research & Therapy (2023)
-
FOXO1 promotes endothelial cell elongation and angiogenesis by up-regulating the phosphorylation of myosin light chain 2
Angiogenesis (2023)
-
Building gut from scratch — progress and update of intestinal tissue engineering
Nature Reviews Gastroenterology & Hepatology (2022)
-
Neural crest cell-derived pericytes act as pro-angiogenic cells in human neocortex development and gliomas
Fluids and Barriers of the CNS (2021)
-
Mitigating the non-specific uptake of immunomagnetic microparticles enables the extraction of endothelium from human fat
Communications Biology (2021)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.