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Regeneration and orthotopic transplantation of a bioartificial lung

Nature Medicine volume 16, pages 927–933 (2010)Cite this article

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Abstract

About 2,000 patients now await a donor lung in the United States. Worldwide, 50 million individuals are living with end-stage lung disease. Creation of a bioartificial lung requires engineering of viable lung architecture enabling ventilation, perfusion and gas exchange. We decellularized lungs by detergent perfusion and yielded scaffolds with acellular vasculature, airways and alveoli. To regenerate gas exchange tissue, we seeded scaffolds with epithelial and endothelial cells. To establish function, we perfused and ventilated cell-seeded constructs in a bioreactor simulating the physiologic environment of developing lung. By day 5, constructs could be perfused with blood and ventilated using physiologic pressures, and they generated gas exchange comparable to that of isolated native lungs. To show in vivo function, we transplanted regenerated lungs into orthotopic position. After transplantation, constructs were perfused by the recipient's circulation and ventilated by means of the recipient's airway and respiratory muscles, and they provided gas exchange in vivo for up to 6 h after extubation.

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Figure 1: Perfusion decellularization of whole rat lungs.
Figure 2: Histology of regenerated lung constructs.
Figure 3: Protein analysis, morphometry and stereology of regenerated lung constructs.
Figure 4: In vitro functional testing of regenerated lung constructs.
Figure 5: Orthotopic transplantation and in vivo function.

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Acknowledgements

We thank Harvard Apparatus Inc. for providing bioreactor components and, in particular, J. Consiglio, R. Zink and T. Beha for technical support. We thank J. Titus for assistance with animal surgeries and video recording. We further thank E. Bassett and D. Hoganson for their help with bioreactor parts and materials, A. Pardo for her technical support with western blots and K. Kulig for assistance with confocal microscopy. Electron microscopy was performed in the Microscopy Core of the Center for Systems Biology/Program in Membrane Biology, which is partially supported by Inflammatory Bowel Disease Grant DK43351 and Boston Area Diabetes and Endocrinology Research Center Award DK57521.This study was supported by a Faculty Development Grant provided by the Department of Surgery, Massachusetts General Hospital and by a Young Clinician Researcher Award granted by the Center for Integration of Medicine and Innovative Technology (CIMIT).

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Authors and Affiliations

  1. Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Harald C Ott, Ben Clippinger, Claudius Conrad, Christian Schuetz, Irina Pomerantseva & Joseph P Vacanti

  2. The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA

    Laertis Ikonomou & Darrell Kotton

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  1. 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. He was responsible for the primary undertaking, completion and supervision of all experiments. B.C. was responsible for cell culture and preparation of cell suspensions. C.S. characterized fetal lung cells. C.C. assisted in animal surgeries. I.P. provided advice on animal protocols and histology techniques. L.I. performed immunohistochemistry. D.K. provided input on developmental aspects and reviewed and edited the manuscript. J.P.V. provided input on tissue engineering aspects and reviewed and edited the manuscript.

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Correspondence to Harald C Ott.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Methods (PDF 703 kb)

Supplementary Video 1

Initiation of dry ventilation of a regenerated lung construct. (MOV 1075 kb)

Supplementary Video 2

Blood perfusion and ventilation of a regenerated lung construct. (MOV 1190 kb)

Supplementary Video 3

Orthotopic transplantation of a regenerated left lung construct. (MOV 3865 kb)

Supplementary Video 4

Fluoroscopy after orthotopic transplantation of a regenerated left lung construct. (MOV 1771 kb)

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Ott, H., Clippinger, B., Conrad, C. et al. Regeneration and orthotopic transplantation of a bioartificial lung. Nat Med 16, 927–933 (2010). https://doi.org/10.1038/nm.2193

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