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Telomerized human microvasculature is functional in vivo

Nature Biotechnology volume 19, pages 219–224 (2001)Cite this article

Abstract

Previously we showed the superior in vitro survival of human telomerase reverse transcriptase (hTERT)-transduced human endothelial cells (EC). Here we show that retroviral-mediated transduction of hTERT in human dermal microvascular EC (HDMEC) results in cell lines that form microvascular structures when subcutaneously implanted in severe combined immunodeficiency (SCID) mice. Anti-human type IV collagen basement membrane immunoreactivity and visualization of enhanced green fluorescent protein (eGFP)-labeled microvessels confirmed the human origin of these capillaries. No human vasculature was observed after implantation of HT1080 fibrosarcoma cells, 293 human embryonic kidney cells, or human skin fibroblasts. Intravascular red fluorescent microspheres injected into host circulation were found within green “telomerized” microvessels, indicating functional murine–human vessel anastamoses. Whereas primary HDMEC-derived vessel density decreased with time, telomerized HDMEC maintained durable vessels six weeks after xenografting. Modulation of implant vessel density by exposure to different angiogenic and angiostatic factors demonstrated the utility of this system for the study of human microvascular remodeling in vivo.

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Figure 1: Characterization of telomerase activity and fluorescence signal of eGFP-labeled primary HDMEC and telomerized cells.
Figure 2: In vitro tubule formation in primary and telomerized HDMEC using 3D Matrigel.
Figure 3: In vivo tubule formation in SCID mice xenografted with HDMEC.
Figure 4: Specificity of HDMEC-GT at forming in vivo tubules in SCID mice.
Figure 5: Telomerized human microvessels communicate with host murine circulatory system.
Figure 6: Effect of pro- and anti-angiogenic factors on HDMEC-GT-derived microvessels in vivo.

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Acknowledgements

Garry Nolan (Stanford University, Stanford, CA) provided the LZRS retroviral vector and the Phoenix packaging cell line. The LZRS-eGFP construct was provided by Helen Deng (Stanford University, Stanford, CA). Endostatin-specific IgG was a gift from Rupert Timpl (Max Planck Institute, Martinsreid, Germany). We thank Michel Prunieras for his advice and many helpful discussions. This work was supported by NIH PO-1 AR44012, the Carl J. Herzog Foundation, and the Dermatology Foundation. J. Yang received a research fellowship from Warner Lambert Consumer Healthcare through the Dermatology Foundation. The work by W.C.M. was performed under the auspices of the U. S. Department of Energy by Lawrence Livermore National Laboratory under Contract No. W-7405-Eng-48. W.C.M. was also supported in part by NIH AI22511. G.S.H. is a Terman Fellow.

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Authors and Affiliations

  1. Department of Dermatology, Stanford University School of Medicine, Stanford, 94305, CA

    Jiwei Yang, Usha Nagavarapu, Kenneth Relloma & G. Scott Herron

  2. The Molecular Medicine Research Institute, Mountain View, 94043, CA

    Jiwei Yang, Usha Nagavarapu, Kenneth Relloma & G. Scott Herron

  3. Universal Imaging Corporation, West Chester, 19380, PA

    Michael D. Sjaastad

  4. Lawrence Livermore National Laboratory, Livermore, 94550, CA

    William C. Moss

  5. Department of Pathology, Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, 21201, MD

    Antonino Passaniti

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Correspondence to G. Scott Herron.

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Yang, J., Nagavarapu, U., Relloma, K. et al. Telomerized human microvasculature is functional in vivo. Nat Biotechnol 19, 219–224 (2001). https://doi.org/10.1038/85655

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