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Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues

Nature Materials volume 11, pages 768–774 (2012)Cite this article

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Abstract

In the absence of perfusable vascular networks, three-dimensional (3D) engineered tissues densely populated with cells quickly develop a necrotic core1. Yet the lack of a general approach to rapidly construct such networks remains a major challenge for 3D tissue culture2,3,4. Here, we printed rigid 3D filament networks of carbohydrate glass, and used them as a cytocompatible sacrificial template in engineered tissues containing living cells to generate cylindrical networks that could be lined with endothelial cells and perfused with blood under high-pressure pulsatile flow. Because this simple vascular casting approach allows independent control of network geometry, endothelialization and extravascular tissue, it is compatible with a wide variety of cell types, synthetic and natural extracellular matrices, and crosslinking strategies. We also demonstrated that the perfused vascular channels sustained the metabolic function of primary rat hepatocytes in engineered tissue constructs that otherwise exhibited suppressed function in their core.

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Figure 1: Carbohydrate-glass material properties and filament-architecture formation.
Figure 2: Monolithic tissue construct containing patterned vascular architectures and living cells.
Figure 3: Demonstrated control over the three key compartments of vascularized solid tissues.
Figure 4: Perfusion of channels sustains cellular metabolic function in the core of thick, densely populated tissue constructs.

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Acknowledgements

We thank the large number of open source and related projects that critically facilitated this work, including Arduino.cc, RepRap.org, MakerBot.com, Replicat.org, MakerGear.com, Ultimachine.com, Hive76.org, Python.org, Hugin.SourceForge.net, ImageMagick.org, Blender.org, Enblend.sourceforge.net, NIH ImageJ, and Fiji.sc. We thank R. J. Vlacich and C. D. Thompson for assistance with precision pneumatic extrusion, A. Dominguez for assistance with red blood cell isolation, and Y-J. Chen for assistance with transduction. This work was supported in part by grants from the US National Institutes of Health (EB00262, EB08396, GM74048), the Penn Center for Engineering Cells and Regeneration, and the American Heart Association-Jon Holden DeHaan Foundation. Individual fellowship support was provided by R. L. Kirschstein National Research Service Awards from NIH (J.S.M., HL099031; K.R.S., DK091007), the National Science Foundation IGERT program (M.T.Y., DGE-0221664), and the American Heart Association (X.Y., 10POST4220014).

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Authors and Affiliations

  1. Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA

    Jordan S. Miller, Michael T. Yang, Brendon M. Baker, Duc-Huy T. Nguyen, Daniel M. Cohen, Esteban Toro, Peter A. Galie, Xiang Yu, Ritika Chaturvedi & Christopher S. Chen

  2. Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA

    Kelly R. Stevens, Alice A. Chen & Sangeeta N. Bhatia

  3. Howard Hughes Medical Institute, Cambridge, Massachusetts 02139, USA

    Sangeeta N. Bhatia

  4. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA

    Sangeeta N. Bhatia

Authors
  1. Jordan S. Miller
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Contributions

J.S.M. and C.S.C. conceived and initiated the project. J.S.M., K.R.S., M.T.Y., B.M.B., D-H.T.N., D.M.C., E.T., A.A.C., P.A.G., X.Y., and R.C. designed and performed experiments. C.S.C. and S.N.B. supervised the project.

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Correspondence to Christopher S. Chen.

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The authors declare no competing financial interests.

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Miller, J., Stevens, K., Yang, M. et al. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nature Mater 11, 768–774 (2012). https://doi.org/10.1038/nmat3357

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