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Nature Biotechnology
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Engineering vascularized skeletal muscle tissue

Nature Biotechnologyvolume 23, pages879–884 (2005)Cite this article

Abstract

One of the major obstacles in engineering thick, complex tissues such as muscle is the need to vascularize the tissuein vitro. Vascularizationin vitro could maintain cell viability during tissue growth, induce structural organization and promote vascularization upon implantation. Here we describe the induction of endothelial vessel networks in engineered skeletal muscle tissue constructs using a three-dimensional multiculture system consisting of myoblasts, embryonic fibroblasts and endothelial cells coseeded on highly porous, biodegradable polymer scaffolds. Analysis of the conditions for induction and stabilization of the vesselsin vitro showed that addition of embryonic fibroblasts increased the levels of vascular endothelial growth factor expression in the construct and promoted formation and stabilization of the endothelial vessels. We studied the survival and vascularization of the engineered muscle implantsin vivo in three different models. Prevascularization improved the vascularization, blood perfusion and survival of the muscle tissue constructs after transplantation.

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Figure 1:In vitro vascularization of engineered skeletal muscle tissue
Figure 2: Quantitative analysis of endothelial vessels in muscle constructs,in vitro.
Figure 3:In vivo analysis of engineered muscle constructs (a) Differentiation of engineered musclein vivo.

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Acknowledgements

The authors thank the MIT division of comparative medicine for excellent assistance in tissue embedding and processing, Adam Kapur for help with data analysis, and Justin S. Golub for help with RT-PCR assays. We would like to thank Joseph Itskovitz-Eldor for assistance and cooperation in conducting this research. This work was supported by National Institutes of Health grants HL60435 (R.L. and S.L.) and EY05318 (P.A.D. and D.C.D.).

Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, Technion, Haifa, 32000, Israel

    Shulamit Levenberg

  2. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Shulamit Levenberg, Mara Macdonald & Robert Langer

  3. Institute for Biomedical Technology, Twente University, Prof. Bronkhorstlaan 10-D, Bilthoven, 3723MB, The Netherlands

    Jeroen Rouwkema & Clemens A van Blitterswijk

  4. Department of Surgery, Brigham and Women's Hospital, 75 Francis St., Boston, 02115, Massachusetts, USA

    Evan S Garfein

  5. Department of Pediatrics, Massachusetts General Hospital, 55 Fruit St., Boston, 02114, Massachusetts, USA

    Daniel S Kohane

  6. The Schepens Eye Research Institute and Department of Ophthalmology, 20 Staniford St., Boston, 02114, Massachusetts, USA

    Diane C Darland & Patricia A D'Amore

  7. Division of Comparative Medicine, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Robert Marini

  8. Department of Molecular Medicine, Children's Hospital 300 Longwood Avenue, 02115, Harvard Medical School, Boston, Massachusetts, USA

    Richard C Mulligan

Authors
  1. Shulamit Levenberg

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  2. Jeroen Rouwkema

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  3. Mara Macdonald

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  4. Evan S Garfein

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  5. Daniel S Kohane

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  6. Diane C Darland

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  7. Robert Marini

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  8. Clemens A van Blitterswijk

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  9. Richard C Mulligan

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  10. Patricia A D'Amore

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  11. Robert Langer

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Corresponding authors

Correspondence toShulamit Levenberg orRobert Langer.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

VEGF and PDGF-B expression in 3D constructs. (PDF 873 kb)

Supplementary Fig. 2

Quantitative analysis of number of endothelial vessels in muscle implants seeded with HUVEC or hESC-derived endothelial cells. (PDF 481 kb)

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Levenberg, S., Rouwkema, J., Macdonald, M.et al. Engineering vascularized skeletal muscle tissue.Nat Biotechnol23, 879–884 (2005). https://doi.org/10.1038/nbt1109

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