Induction, differentiation, and remodeling of blood vessels after transplantation of Bcl-2-transduced endothelial cells.
Implants of collagen-fibronectin gels containing Bcl-2-transduced human umbilical vein endothelial cells (Bcl-2-HUVECs) induce the formation of human endothelial cell (EC)/murine vascular smooth muscle cell (VSMC) chimeric vessels in immunodeficient mice. Microfil casting of the vasculature 60 d after implantation reveals highly branched microvascular networks within the implants that connect with and induce remodeling of conduit vessels arising from the abdominal wall circulation. Approximately 85% of vessels within the implants are lined by Bcl-2-positive human ECs expressing VEGFR1, VEGFR2, and Tie-2, but not integrin alpha(v)beta(3). The human ECs are seated on a well formed human laminin/collagen IV-positive basement membrane, and are surrounded by mouse VSMCs expressing SM-alpha actin, SM myosin, SM22alpha, and calponin, all markers of contractile function. Transmission electron microscopy identified well formed EC-EC junctions, chimeric arterioles with concentric layers of contractile VSMC, chimeric capillaries surrounded by pericytes, and chimeric venules. Bcl-2-HUVEC-lined vessels retain 70-kDa FITC-dextran, but not 3-kDa dextran; local histamine rapidly induces leak of 70-kDa FITC-dextran or India ink. As in skin, TNF induces E-selectin and vascular cell adhesion molecule 1 only on venular ECs, whereas intercellular adhesion molecule-1 is up-regulated on all human ECs. Bcl-2-HUVEC implants are able to engraft within and increase perfusion of ischemic mouse gastrocnemius muscle after femoral artery ligation. These studies show that cultured Bcl-2-HUVECs can differentiate into arterial, venular, and capillary-like ECs when implanted in vivo, and induce arteriogenic remodeling of the local mouse vessels. Our results support the utility of differentiated EC transplantation to treat tissue ischemia.
Enis
David R
DR
Interdepartmental Program in Vascular Biology and Transplantation, Boyer Center for Molecular Medicine, and Department of Pathology, Yale University School of Medicine, New Haven, CT 06510, USA.
Shepherd
Benjamin R
BR
Wang
Yinong
Y
Qasim
Asif
A
Shanahan
Catherine M
CM
Weissberg
Peter L
PL
Kashgarian
Michael
M
Pober
Jordan S
JS
Schechner
Jeffrey S
JS
D000818:Animals; D002199:Capillary Permeability; D002454:Cell Differentiation; D042783:Endothelial Cells / Q000637:transplantation*; D006614:Hindlimb / Q000098:blood supply; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007511:Ischemia / Q000628:therapy*; D051379:Mice; D008854:Microscopy, Electron; D018919:Neovascularization, Physiologic; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D019253:Proto-Oncogene Proteins c-bcl-2; D014409:Tumor Necrosis Factor-alpha; D006632:Histamine