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Spheroid-based engineering of a human vasculature in mice

Nature Methods volume 5, pages 439–445 (2008)Cite this article

Abstract

The complexity of the angiogenic cascade limits cellular approaches to studying angiogenic endothelial cells (ECs). In turn, in vivo assays do not allow the analysis of the distinct cellular behavior of ECs during angiogenesis. Here we show that ECs can be grafted as spheroids into a matrix to give rise to a complex three-dimensional network of human neovessels in mice. The grafted vasculature matures and is connected to the mouse circulation. The assay is highly versatile and facilitates numerous applications including studies of the effects of different cytokines on angiogenesis. Modifications make it possible to study human lymphangiogenic processes in vivo. EC spheroids can also be coimplanted with other cell types for tissue engineering purposes.

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Figure 1: Characteristics of human neovasculature originating from transplanted EC spheroids.
Figure 2: Vascularization originating from luciferase-expressing HUVEC spheroids in a Matrigel-fibrin matrix containing VEGF and FGF-2 subcutaneously injected into SCID mice.
Figure 3: Analysis of VEGF- and FGF-2–induced angiogenesis.
Figure 4: Validation of antiangiogenic drugs using the VEGFR inhibitor PTK/ZK.
Figure 5: Growth of lymphatic vessels from podoplanin-positive LECs grafted as spheroids in a Matrigel-fibrin matrix.

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Acknowledgements

This work was supported by grants from the German Research Council (Deutsche Forschungsgemeinschaft, AU83/10-1), the European Union (LSHG-CT-2004-503573), the Austrian Science Fund (FWF S9404-B11), the State of Baden-Württemberg Kompetenznetzwerk Biomaterialien and the Fördergesellschaft der Klinik für Tumorbiologie Freiburg.

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Author notes

  1. Abdullah Alajati, Holger Weber, Thomas Korff & Cynthia Obodozie

    Present address: Present addresses: Friedrich-Miescher Institute, Maulbeerstrasse 66, CH-4058 Basel, Switzerland (A.A.); ProQinase GmbH, Breisacher Str. 117, D-79108 Freiburg, Germany (H.W., C.O.); Department of Physiology, University of Heidelberg, Im Neuenheimer Feld 581, D-69120 Heidelberg, Germany (T.K.).,

  2. Abdullah Alajati, Anna M Laib and Holger Weber: These authors contributed equally to this study.

Authors and Affiliations

  1. Joint Research Division Vascular Biology of the Medical Faculty Mannheim (CBTM), University of Heidelberg, Ludolph-Krehl-Str. 11-14, D-68167 Mannheim, Germany, and the German Cancer Research Center (DKFZ), Im Neuenheimer Feld 581, D-69120 Heidelberg, Germany.,

    Abdullah Alajati, Anna M Laib, Anja M Boos, Arne Bartol, Sven Christian, Mélanie Héroult & Hellmut G Augustin

  2. Department of Vascular Biology and Angiogenesis Research, Tumor Biology Center, Breisacher Str. 117, Freiburg, D-79108, Germany

    Abdullah Alajati, Holger Weber, Anja M Boos, Kristian Ikenberg, Thomas Korff, Cynthia Obodozie, Sven Christian, Mélanie Héroult & Hellmut G Augustin

  3. Department of Plastic and Hand Surgery, University of Freiburg Medical Center, Hugstetter Str. 55, Freiburg, D-79106, Germany

    Abdullah Alajati, Günter Finkenzeller & G Björn Stark

  4. Electron Microscopy Group, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, Heidelberg, D-69120, Germany

    Hanswalter Zentgraf

  5. ProQinase GmbH, Breisacher Str. 117, Freiburg, D-79108, Germany

    Ralph Graeser

Authors
  1. Abdullah Alajati
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Contributions

A.A., A.M.L., H.W., S.C., M.H., G.B.S. and H.G.A. conceived and designed the experiments. A.A., A.M.L., H.W., A.M.B., A.B., K.I., T.K., H.Z., C.O., R.G., G.F. and M.H. performed experiments. A.A., A.M.L., H.W., M.H., S.C. and H.G.A. contributed to data analysis. A.A., A.M.L., H.W., M.H. and H.G.A. contributed to the writing of the manuscript. All authors discussed and commented on the manuscript.

Corresponding author

Correspondence to Hellmut G Augustin.

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Supplementary Figures 1–6, Supplementary Methods (PDF 3008 kb)

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Alajati, A., Laib, A., Weber, H. et al. Spheroid-based engineering of a human vasculature in mice. Nat Methods 5, 439–445 (2008). https://doi.org/10.1038/nmeth.1198

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