Date of Award
Open Access Thesis
Medical Doctor (MD)
Frank J Giordano
This study explored the possibility that upregulation of Hypoxia Inducible Factor-1 (Hif-1)-responsive genes in Human Umbilical Vein Endothelial Cells (HUVEC) would promote and stabilize HUVEC formation into inchoate vascular beds within artificial collagen gels. This experiment was designed to explore the above possibility by sub-cloning Hif-1α, the related chimeric construct Hif-1α/VP16, and the marker gene dsRed into retroviral expression vectors, producing retroviral vectors containing these genes, and stably transducing HUVEC using these retroviruses. Transduced HUVEC were to be observed in cell culture as well as after implantation into artificial collagen gels that have previously supported vascular bed formation by HUVEC. Our results show, preliminarily, that HUVEC transduced with Hif-1α/VP16 go into cell-cycle arrest. Attempts to transduce HUVEC with Hif-1α failed to achieve high enough transduction efficiency to determine the cells angiogenic potential. This study concluded that more experiments need to be conducted to better characterize the effects of hypoxia-responsive gene upregulation in controlling HUVEC angiogenesis and cell-cycle signaling and that straightforward transduction of HUVEC by Hif-1α/VP16 is probably not sufficient, in itself, to induce in vitro vascular bed formation.
Schonberger, Robert Brian, "Upregulation of Hypoxia-Inducible Genes in Endothelial Cells to Create Artificial Vasculature" (2006). Yale Medicine Thesis Digital Library. 289.