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Blood, 15 February 2004, Vol. 103, No. 4, pp. 1325-1332.
Prepublished online as a Blood First Edition Paper on October 16, 2003; DOI 10.1182/blood-2003-03-0799.
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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Functional endothelial cells derived from rhesus monkey embryonic stem cells
Dan S. Kaufman,
Rachel L. Lewis,
Eric T. Hanson,
Robert Auerbach,
Johanna Plendl, and
James A. Thomson
From the Wisconsin National Primate Research Center, the Laboratory of Developmental Biology, and the Department of Anatomy, University of Wisconsin, Madison; and the Freie Universität Berlin, Fachbereich Veterinärmedizin, Institut für Veterinär-Anatomie, Germany.
We have used rhesus monkey embryonic stem (ES) cells to study endothelial cell development. Rhesus ES cells (R366.4 cell line) exposed to medium containing vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF), and epidermal growth factor (EGF) assumed a relatively uniform endothelial cell morphology and could be propagated and expanded with a consistent phenotype and normal karyotype. When placed in Matrigel, these rhesus ES cell–derived endothelial cells (RESDECs) formed capillary-like structures characteristic of endothelial cells. Immunohistochemical and flow cytometric analysis of RESDECs showed that they take up acetylated low-density lipoprotein (LDL), express CD146, von Willebrand factor, and the integrin  v 3, and bind the lectin ulex europaeus agglutinin-1. These cells also express the VEGF receptor Flk-1 and secrete VEGF. When introduced in a Matrigel plug implanted subcutaneously in mice, RESDECs formed intact vessels and recruited new endothelial cell growth. In vivo function was demonstrated by coinjection of RESDECs with murine tumor cells subcutaneously into immunocompromised adult mice. RESDECs injected alone did not form measurable tumors. Tumor cells grew more rapidly and had increased vascularization when coinjected with the RESDECs. Immunohistochemical staining demonstrated that the RESDECs participated in forming the tumor neovasculature. RESDECs provide a novel means to examine the mechanisms of endothelial cell development, and may open up new therapeutic strategies.
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