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Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-functionFPS/FES proto-oncogene

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Summary

We report on the isolation and propagation of endothelial cells from the mouse embryonic yolk sac, the earliest site of blood vessel development, and on the advantages of a hypervascular transgenic mouse source of these cells. These transgenic mice express multiple copies of an activated allele of the humanfps/fes proto-oncogene and display hypervascularity progressing to multifocal hemangiomas. This phenotype suggested a role of thefps/fes proto-oncogene in vasculogenesis and angiogenesis and led us to investigate the growth characteristics of yolk-sac-derived endothelial cells from transgenicfps/fes embryos. We have established eight independent cell clones from a mixture of transgenic and control yolk sacs from Day 12 embryos. Southern blot hybridization analysis showed all eight clones to be derived from transgenic cells suggesting a growth advantage of cells carrying the activatedfps/fes gene. A cell line, Clone 166 (C166), established from one of these clones, was more fully characterized. C166 exhibits normal endothelial characteristics, such as rearrangement into tubelike structures when placed on Matrigel, expression of angiotensin converting enzyme, retention of cobblestone morphology at confluence, and the presence of cell surface receptors for acetylated low density lipoprotein. The cells constitutively express murine endothelial cell adhesion molecule VCAM-1 and the vascular addressin identified by antibody MECA-99. As expected, the cell line expresses high levels of the cytoplasmic protein-tyrosine kinase encoded by thefps/fes proto-oncogene. The clone we have described as well as other endothelial cell lines that we have established from the mouse embryonic yolk sac should prove useful for the study of endothelial cell differentiation and for the determination of the mechanisms underlying the establishment of organ-specific endothelial cell heterogeneity.

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Author notes

  1. Shur-Jen Wang

    Present address: Center for Developmental Biology, University of Wisconsin, 1117 W. Johnson Street, 53706, Madison, Wisconsin

  2. Peter Greer

    Present address: Cancer Research Laboratories, Queen's University, Kingston, Ontario, Canada

Authors and Affiliations

  1. Center for Developmental Biology, University of Wisconsin, 1117 W. Johnson Street, 53706, Madison, Wisconsin

    Robert Auerbach

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  1. Shur-Jen Wang
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  2. Peter Greer
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  3. Robert Auerbach
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Wang, SJ., Greer, P. & Auerbach, R. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-functionFPS/FES proto-oncogene. In Vitro Cell.Dev.Biol.-Animal 32, 292–299 (1996). https://doi.org/10.1007/BF02723062

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  • DOI: https://doi.org/10.1007/BF02723062

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