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Embryonic Stem Cell Differentiation and the Vascular Lineage

  • Victoria L. Bautch
Part of the Methods in Molecular Biology™ book series (MIMB, volume 185)

Abstract

The ability of mouse embryonic stem (ES) cells to undergo differentiation in vitro complements their ability to contribute to numerous tissues in vivo and provides a unique model system for aspects of early mammalian development. ES cells are differentiated in two major ways: (i) unmanipulated differentiation involves the removal of differentiation inhibitory factors, allowing the ES cells to undergo a programmed differentiation to form multiple cell types that provide developmental cues to each other; and (ii) manipulated differentiation begins with the removal of differentiation inhibitory factors, but at some point the cells are usually disaggregated and cultured with specific added factors to purify or to increase the proportion of cells that acquire a particular developmental fate. Examples of both kinds of differentiation are found in this volume. The protocols provided here are for unmanipulated differentiation, which reproducibly results in the development of a primitive vasculature. Endothelial cells typically comprise 15–20% of the differentiated ES cells.

Keywords

Embryonic Stem Cell Tissue Culture Dish Vascular Development Cell Clump Embryonic Stem Cell Differentiation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Doetschman, T. C., Eistetter, H., Katz, M., Schmidt, W., and 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. Morph. 87, 27–45.PubMedGoogle Scholar
  2. 2.
    Risau, W., Sariola, H., Zerwes, H. G., Sasse, J., Ekblom, P., Kemler, R., and Doetschman, T. (1988) Vasculogenesis and angiogenesis in embryonic-stem-cell-derived embryoid bodies. Development 102, 471–478.PubMedGoogle Scholar
  3. 3.
    Schmitt, R. M., Bruyns, E., and Snodgrass, H. R. (1991) Hematopoietic development of embryonic stem cells in vitro: cytokine and receptor gene expression. Genes Dev. 5, 728–740.PubMedCrossRefGoogle Scholar
  4. 4.
    Wang, R., Clark, R., and Bautch, V. L. (1992) Embryonic stem cell-derived cystic embryoid bodies form vascular channels: an in vitro model of blood vessel development. Development 114, 303–316.PubMedGoogle Scholar
  5. 5.
    Bautch, V. L., Stanford, W. L., Rapoport, R., Russell, S., Byrum, R. S., and Futch, T. A. (1996) Blood island formation in attached cultures of murine embryonic stem cells. Dev. Dyn. 205, 1–12.PubMedCrossRefGoogle Scholar
  6. 6.
    Inamdar, M., Koch, T., Rapoport, R., Dixon, J. T., Probolus, J. A., Cram, E., and Bautch, V. L. (1997) Yolk sac-derived murine macrophage cell line has a counterpart during ES cell differentiation. Dev. Dyn. 210, 487–497.PubMedCrossRefGoogle Scholar
  7. 7.
    Bautch, V. L., Redick, S. D., Scalia, A., Harmaty, M., Carmeliet, P., and Rapoport, R. (2000) Characterization of the vasculogenic block in the absence of vascular endothelial growth factor-A. Blood 95, 1979–1987.PubMedGoogle Scholar
  8. 8.
    Albelda, S. M., Muller, W. A., Buck, C. A., and Newman, P. J. (1991) Molecular and cellular properties of PECAM-1 (endoCAM/CD31): a novel vascular cell-cell adhesion molecule. J. Cell Biol. 114, 1059–1068.PubMedCrossRefGoogle Scholar
  9. 9.
    Piali, L., Albelda, S. M., Baldwin, H. S., Hammel, P., Ginsler, R. H., and Imhof, B. A. (1993) Murine platelet endothelial cell adhesion molecule (PECAM-1)/CD31 modulates β2 integrins on lymphocyte-activated killer cells. Eur. J. Immunol. 23, 2464–2471.PubMedCrossRefGoogle Scholar
  10. 10.
    Baldwin, H. S., Shen, H. M., Yan, H. C., DeLisser, H. M., Chung, A., Mickanin, C., et al. (1994) Platelet endothelial cell adhesion molecule-1 (PECAM/CD31): alternatively spliced, functionally distinct isoforms expressed during mammalian cardiovascular development. Development 120, 2539–2553.PubMedGoogle Scholar
  11. 11.
    Vittet, D., Prandini, M.-H., Berthier, R., Schweitzer, A., Martin-Sisteron, H., Uzan, G., and Dejana, E. (1996) Embryonic stem cells differentiate in vitro to endothelial cells through successive maturation steps. Blood 88, 3424–3431.PubMedGoogle Scholar
  12. 12.
    Shalaby, F., Ho, J., Stanford, W. L., Fischer, K.-D., Schuh, A. C., Schwartz, L., et al. (1997) A requirement for Flk-1 in primitive and definitive hematopoiesis and vasculogenesis. Cell 89, 981–990.PubMedCrossRefGoogle Scholar
  13. 13.
    Schuh, A. C., Faloon, P., Hu, Q.-L., Bhimani, M., and Choi, K. (1999) In vitro hematopoietic and endothelial potential of flk-1 ―/― embryonic stem cells and embryos. Proc. Natl. Acad. Sci. USA 96, 2159–2164.PubMedCrossRefGoogle Scholar
  14. 14.
    Fong, G.-H., Zhang, L., Bryce, D. M., and Peng, J. (1999) Increased hemangioblast commitment, not vascular disorganization, is the primary defect in flt-1 knock-out mice. Development 126, 3015–3025.PubMedGoogle Scholar
  15. 15.
    Hidaka, M., Stanford, W. L., and Bernstein, A. (1999) Conditional requirement for the Flk-1 receptor in the in vitro generation of early hematopoietic cells. Proc. Natl. Acad. Sci. USA 96, 7370–7375.PubMedCrossRefGoogle Scholar
  16. 16.
    Stanford, W. L., Caruana, G., Vallis, K. A., Inamdar, M., Hidaka, M., Bautch, V. L., and Bernstein, A. (1998) Expression trapping: identification of novel genes expressed in hematopoietic and endothelial lineages by gene trapping in ES cells. Blood 92, 4622–4631.PubMedGoogle Scholar
  17. 17.
    Kearney, J.B., Ambler, C.A., Monaco, K., Johnson, N., Rapoport, R., and Bautch, V. L. (2000) The VEGF receptor flt-1 negatively regulates blood vessel formation by modulating endothelial cell division. (Submitted).Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Victoria L. Bautch
    • 1
  1. 1.Department of BiologyThe University of North Carolina at Chapel HillChapel Hill

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