caa a22 4500 477081908 CHVBK 20180405111452.0 cr unu---uuuuu 170330e19960501xx s 000 0 eng 10.1007/BF02723062 doi (NATIONALLICENCE)springer-10.1007/BF02723062 Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function FPS/FES proto-oncogene [Elektronische Daten] [Shur-Jen Wang, Peter Greer, Robert Auerbach] 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. Society for In Vitro Biology, 1996 cell culture nationallicence flow cytometry nationallicence yolk sac endothelium nationallicence blood vessel development nationallicence Wang Shur-Jen Center for Developmental Biology, University of Wisconsin, 1117 W. Johnson Street, 53706, Madison, Wisconsin aut Greer Peter Cancer Research Laboratories, Queen's University, Kingston, Ontario, Canada aut Auerbach Robert Center for Developmental Biology, University of Wisconsin, 1117 W. Johnson Street, 53706, Madison, Wisconsin aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/5(1996-05-01), 292-299 1071-2690 32:5<292 1996 32 11626 https://doi.org/10.1007/BF02723062 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02723062 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Shur-Jen Center for Developmental Biology, University of Wisconsin, 1117 W. Johnson Street, 53706, Madison, Wisconsin aut NATIONALLICENCE 700 1- Greer Peter Cancer Research Laboratories, Queen's University, Kingston, Ontario, Canada aut NATIONALLICENCE 700 1- Auerbach Robert Center for Developmental Biology, University of Wisconsin, 1117 W. Johnson Street, 53706, Madison, Wisconsin aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/5(1996-05-01), 292-299 1071-2690 32:5<292 1996 32 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer