Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues (original) (raw)

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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References

  1. Radisic, M. et al. Medium perfusion enables engineering of compact and contractile cardiac tissue. Am. J. Physiol. Heart Circ. Physiol. 286, H507–H516 (2004).
    Article CAS Google Scholar
  2. Langer, R. & Vacanti, J. P. Tissue engineering. Science 260, 920–926 (1993).
    Article CAS Google Scholar
  3. Griffith, L. G. & Swartz, M. A. Capturing complex 3D tissue physiology in vitro. Nature Rev. Mol. Cell Biol. 7, 211–224 (2006).
    Article CAS Google Scholar
  4. Vunjak-Novakovic, G. & Scadden, D. T. Biomimetic platforms for human stem cell research. Cell Stem Cell 8, 252–261 (2011).
    Article CAS Google Scholar
  5. Schmidt-Nielsen, K. Scaling in biology: The consequences of size. J. Exp. Zool. 194, 287–307 (1975).
    Article CAS Google Scholar
  6. Golden, A. P. & Tien, J. Fabrication of microfluidic hydrogels using molded gelatin as a sacrificial element. Lab Chip 7, 720–725 (2007).
    Article CAS Google Scholar
  7. Ling, Y. et al. A cell-laden microfluidic hydrogel. Lab Chip 7, 756–762 (2007).
    Article CAS Google Scholar
  8. Choi, N. W. et al. Microfluidic scaffolds for tissue engineering. Nature Mater. 6, 908–915 (2007).
    Article CAS Google Scholar
  9. Cuchiara, M. P., Allen, A. C. B., Chen, T. M., Miller, J. S. & West, J. L. Multilayer microfluidic PEGDA hydrogels. Biomaterials 31, 5491–5497 (2010).
    Article CAS Google Scholar
  10. Visconti, R. P. et al. Towards organ printing: Engineering an intra-organ branched vascular tree. Exp. Opin. Biol. Therapy 10, 409–420 (2010).
    Article Google Scholar
  11. Therriault, D., White, S. R. & Lewis, J. A. Chaotic mixing in three-dimensional microvascular networks fabricated by direct-write assembly. Nature Mater. 2, 265–271 (2003).
    Article CAS Google Scholar
  12. Bellan, L. M. et al. Fabrication of an artificial 3-dimensional vascular network using sacrificial sugar structures. Soft Matter 5, 1354–1357 (2009).
    Article CAS Google Scholar
  13. Wu, W. et al. Direct-write assembly of biomimetic microvascular networks for efficient fluid transport. Soft Matter 6, 739–742 (2010).
    Article CAS Google Scholar
  14. Reinheimer, M. A., Mussati, S. & Scenna, N. J. Influence of product composition and operating conditions on the unsteady behavior of hard candy cooling process. J. Food Eng. 101, 409–416 (2010).
    Article Google Scholar
  15. Lutolf, M. P. & Hubbell, J. A. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nature Biotechnol. 23, 47–55 (2005).
    Article CAS Google Scholar
  16. Tibbitt, M. W. & Anseth, K. S. Hydrogels as extracellular matrix mimics for 3D cell culture. Biotechnol. Bioeng. 103, 655–663 (2009).
    Article CAS Google Scholar
  17. Heiber, J., Clemens, F., Graule, T. & Hülsenberg, D. Thermoplastic extrusion to highly-loaded thin green fibres containing Pb(Zr,Ti)O3 . Adv. Eng. Mater. 7, 404–408 (2005).
    Article CAS Google Scholar
  18. Miller, J. S. et al. Bioactive hydrogels made from step-growth derived PEG-peptide macromers. Biomaterials 31, 3736–3743 (2010).
    Article CAS Google Scholar
  19. Moon, J. J., Hahn, M. S., Kim, I., Nsiah, B. A. & West, J. L. Micropatterning of poly(ethylene glycol) diacrylate hydrogels with biomolecules to regulate and guide endothelial morphogenesis. Tissue Eng. Part A 15, 579–585 (2009).
    Article CAS Google Scholar
  20. DeLong, S. A., Moon, J. J. & West, J. L. Covalently immobilized gradients of bFGF on hydrogel scaffolds for directed cell migration. Biomaterials 26, 3227–3234 (2005).
    Article CAS Google Scholar
  21. Stefonek, T. J. & Masters, K. S. Immobilized gradients of epidermal growth factor promote accelerated and directed keratinocyte migration. Wound Repair Regen. 15, 847–855 (2007).
    Article Google Scholar
  22. Leslie-Barbick, J. E., Moon, J. J. & West, J. L. Covalently-immobilized vascular endothelial growth factor promotes endothelial cell tubulogenesis in poly(ethylene glycol) diacrylate hydrogels. J. Biomater. Sci. Poly. Edn 20, 1763–1779 (2009).
    Article CAS Google Scholar
  23. Sundararaghavan, H. G., Monteiro, G. A., Firestein, B. L. & Shreiber, D. I. Neurite growth in 3D collagen gels with gradients of mechanical properties. Biotechnol. Bioeng. 102, 632–643 (2009).
    Article CAS Google Scholar
  24. Nemir, S., Hayenga, H. N. & West, J. L. PEGDA hydrogels with patterned elasticity: Novel tools for the study of cell response to substrate rigidity. Biotechnol. Bioeng. 105, 636–644 (2010).
    Article CAS Google Scholar
  25. Tsang, V. L. et al. Fabrication of 3D hepatic tissues by additive photopatterning of cellular hydrogels. FASEB J. 21, 790–801 (2007).
    Article CAS Google Scholar
  26. Thorsen, T., Maerkl, S. J. & Quake, S. R. Microfluidic large-scale integration. Science 298, 580–584 (2002).
    Article CAS Google Scholar
  27. Atencia, J. & Beebe, D. J. Controlled microfluidic interfaces. Nature 437, 648–655 (2005).
    Article CAS Google Scholar

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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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  2. Kelly R. Stevens
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  3. Michael T. Yang
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  4. Brendon M. Baker
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  5. Duc-Huy T. Nguyen
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  6. Daniel M. Cohen
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  7. Esteban Toro
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  8. Alice A. Chen
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  9. Peter A. Galie
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  10. Xiang Yu
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  11. Ritika Chaturvedi
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  12. Sangeeta N. Bhatia
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  13. Christopher S. Chen
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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 toChristopher 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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