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Engineering pulmonary vasculature in decellularized rat and human lungs

Nature Biotechnology volume 33, pages 1097–1102 (2015)Cite this article

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Abstract

Bioengineered lungs produced from patient-derived cells may one day provide an alternative to donor lungs for transplantation therapy. Here we report the regeneration of functional pulmonary vasculature by repopulating the vascular compartment of decellularized rat and human lung scaffolds with human cells, including endothelial and perivascular cells derived from induced pluripotent stem cells. We describe improved methods for delivering cells into the lung scaffold and for maturing newly formed endothelium through co-seeding of endothelial and perivascular cells and a two-phase culture protocol. Using these methods we achieved ∼75% endothelial coverage in the rat lung scaffold relative to that of native lung. The regenerated endothelium showed reduced vascular resistance and improved barrier function over the course of in vitro culture and remained patent for 3 days after orthotopic transplantation in rats. Finally, we scaled our approach to the human lung lobe and achieved efficient cell delivery, maintenance of cell viability and establishment of perfusable vascular lumens.

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Figure 1: Endothelial delivery, vascular maturation and functional assessment of regenerated lungs during in vitro culture.
Figure 2: Endothelial and perivascular cell differentiation from hiPSCs.
Figure 3: Pulmonary vascular regeneration with hiPSC-ECs and hiPSC-PCs.
Figure 4: Human lung regeneration using hiPSC-ECs and hiPSC-PCs.

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Acknowledgements

This study was supported by the United Therapeutics Corporation and the National Institutes of Health (NIH) Director's New Innovator Award (DP2-OD008749-01). The authors thank the HSCI/MGH Flow Cytometry Core Facility for analysis and purification of hiPSC differentiation, the MGH Center for Skeletal Research Core (NIH P30 AR066261) for histological processing and MGH Wellman Center for Photomedicine for transmission electron microscopy.

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  1. hott@mgh.harvard.edu

Authors and Affiliations

  1. Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA

    Xi Ren, Philipp T Moser, Sarah E Gilpin, Tatsuya Okamoto, Tong Wu, Luis F Tapias, Francois E Mercier, Linjie Xiong, Raja Ghawi, David T Scadden & Harald C Ott

  2. Harvard Medical School, Boston, Massachusetts, USA

    Xi Ren, Philipp T Moser, Sarah E Gilpin, Tatsuya Okamoto, Tong Wu, Luis F Tapias, Francois E Mercier, Linjie Xiong, Raja Ghawi, David T Scadden, Douglas J Mathisen & Harald C Ott

  3. Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital, Boston, Massachusetts, USA

    Luis F Tapias, Douglas J Mathisen & Harald C Ott

  4. Harvard College, Cambridge, Massachusetts, USA

    Raja Ghawi

  5. Harvard Stem Cell Institute, Cambridge, Massachusetts, USA

    David T Scadden & Harald C Ott

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Contributions

H.C.O. conceived, designed and oversaw all of the studies, collection of results, interpretation of the data and writing of the manuscript and was responsible for the primary undertaking, completion and supervision of all experiments. X.R. designed the studies, conducted cell culture, animal surgeries, decellularization, whole-organ culture, histological and functional analysis, and prepared the manuscript. P.T.M. assisted in animal surgeries, histological analysis and figure preparation. S.E.G. and L.F.T. conducted human lung decellularization, and assisted in human lung recellularization. T.O. and L.X. performed orthotopic lung transplantation. T.W. performed perivascular cell characterization and labeling. F.E.M. designed and constructed the lentiviral vector. R.G. assisted in histological analysis. D.T.S. and D.J.M. provided input on experimental design and edited the manuscript.

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Competing interests

H.C.O. is founder and stockholder of IVIVA Medical Inc. This relationship did not affect the content or conclusions contained in this manuscript.

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Ren, X., Moser, P., Gilpin, S. et al. Engineering pulmonary vasculature in decellularized rat and human lungs. Nat Biotechnol 33, 1097–1102 (2015). https://doi.org/10.1038/nbt.3354

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