In vivo formation of complex microvessels lined by human endothelial cells in an immunodeficient mouse
- Jeffrey S. Schechner*,†,‡,
- Anjali K. Nath*,†,
- Lian Zheng§,
- Martin S. Kluger*,†,
- Christopher C. W. Hughes¶,‖,
- M. Rocio Sierra-Honigmann¶,
- Marc I. Lorber**,
- George Tellides**,
- Michael Kashgarian¶,
- Alfred L. M. Bothwell§, and
- Jordan S. Pober*,†,§,¶
- *Interdepartmental Program in Vascular Biology and Transplantation, Department of †Dermatology, §Section of Immunobiology, and Departments of ¶Pathology and **Surgery, Yale University School of Medicine, New Haven, CT 06536
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Communicated by Vincent T. Marchesi, Yale University School of Medicine, New Haven, CT (received for review February 23, 2000)
Abstract
We have identified conditions for forming cultured human umbilical vein endothelial cells (HUVEC) into tubes within a three-dimensional gel that on implantation into immunoincompetent mice undergo remodeling into complex microvessels lined by human endothelium. HUVEC suspended in mixed collagen/fibronectin gels organize into cords with early lumena by 24 h and then apoptose. Twenty-hour constructs, s.c. implanted in immunodeficient mice, display HUVEC-lined thin-walled microvessels within the gel 31 days after implantation. Retroviral-mediated overexpression of a caspase-resistant Bcl-2 protein delays HUVEC apoptosis in vitro for over 7 days. Bcl-2-transduced HUVEC produce an increased density of HUVEC-lined perfused microvessels in vivo compared with untransduced or control-transduced HUVEC. Remarkably, Bcl-2- but not control-transduced HUVEC recruit an ingrowth of perivascular smooth-muscle α-actin-expressing mouse cells at 31 days, which organize by 60 days into HUVEC-lined multilayered structures resembling true microvessels. This system provides an in vivo model for dissecting mechanisms of microvascular remodeling by using genetically modified endothelium. Incorporation of such human endothelial-lined microvessels into engineered synthetic skin may improve graft viability, especially in recipients with impaired angiogenesis.
Footnotes
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↵ ‡ To whom reprint requests should be addressed at: Department of Dermatology, Yale University School of Medicine, P.O. Box 208059, New Haven, CT 06520-8059. E-mail: Jeffrey.Schechner{at}yale.edu.
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↵ ‖ Present address: Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697.
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Article published online before print: Proc. Natl. Acad. Sci. USA, 10.1073/pnas.150242297.
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Article and publication date are at www.pnas.org/cgi/doi/10.1073/pnas.150242297
- Abbreviations:
- EC,
- endothelial cells;
- 3D,
- three-dimensional;
- HUVEC,
- human umbilical-vein endothelial cells;
- UEA-1,
- Ulex europaeus agglutinin-1;
- SCID,
- severe combined immunodeficient;
- EGFP,
- enhanced green fluorescent protein
- Copyright © The National Academy of Sciences
