Induction, Differentiation, and Remodeling of Blood Vessels after Transplantation of Bcl-2-Transduced Endothelial Cells

• Published in: Proceedings of the National Academy of Sciences - PNAS 2005-01, Vol.102 (2), p.425-430
• Main Authors: Enis, David R., Shepherd, Benjamin R., Wang, Yinong, Qasim, Asif, Shanahan, Catherine M., Weissberg, Peter L., Kashgarian, Michael, Pober, Jordan S., Schechner, Jeffrey S., Marchesi, Vincent T.
• Format: Article
• Language: English
• Subjects: Abdomen; Animals; Biological Sciences; Blood vessels; Capillaries; Capillary Permeability; Cell Differentiation; Cells; Endothelial Cells - transplantation; Gels; Hindlimb - blood supply; Histamine - pharmacology; Histamines; Humans; Immunohistochemistry; Implants; Ischemia - therapy; Membranes; Mice; Microscopy, Electron; Microvessels; Neovascularization, Physiologic; Perfusion; Proto-Oncogene Proteins c-bcl-2 - genetics; Rodents; Transduction, Genetic; Transmission electron microscopy; Transplants & implants; Tumor Necrosis Factor-alpha - pharmacology
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0408357102

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 αvβ3. The human ECs are seated on a well formed human laminin/collagen IV-positive basement membrane, and are surrounded by mouse VSMCs expressing SM-α actin, SM myosin, SM22α, 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.