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Orthotopic mouse lung transplantation as experimental methodology to study transplant and tumor biology

Abstract

Unlike transplantation of other solid organs, vascularized mouse lung transplantation has only recently been developed. In this protocol, we describe a detailed method for performing a vascularized and aerated mouse orthotopic lung transplant, which to date represents the most physiological mouse model of lung transplantation. The procedure is divided into two separate portions consisting of donor harvest followed by implantation using the cuff technique for bronchovascular anastomoses. After a training period spanning several months, the procedure can be successfully mastered and, in experienced hands, requires 90 min to perform. After an initial learning curve, perioperative survival is close to 100%. As the donor hematopoietic cells in the transplanted lung are replaced by those of the host over time, thereby creating a 'chimeric lung,' this model represents a novel research tool for the study of transplantation biology as well as other disease processes, such as malignancies.

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Figure 1: Replacement of hematopoietic cells in the transplanted lung.
Figure 2: Comparison of tumor burden between transplanted and native left lungs.
Figure 3: General principles of cuff technique for establishing microvascular anastomoses.
Figure 4: Custom-made clamp for stabilization of cuffs.
Figure 5: Creation of cuffs.
Figure 6: Donor procedure.
Figure 7: Recipient procedure.

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Acknowledgements

A.S.K. is supported by (i) the American Association for Thoracic Surgery Norman Shumway Research Scholarship; (ii) American Cancer Society Internal Research Grant through the Alvin J Siteman Cancer Center; (iii) Melanoma Research Foundation; and (iv) a grant jointly sponsored by The National Cancer Institute and The Thoracic Surgery Foundation for Research and Education (1K08CA131097). D.K. is supported by a grant jointly sponsored by The National Heart, Lung and Blood Institute and The Thoracic Surgery Foundation for Research and Education (1K08HL083983-01). We thank Arlene Aligori for the medical illustrations.

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Correspondence to Alexander S Krupnick.

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Krupnick, A., Lin, X., Li, W. et al. Orthotopic mouse lung transplantation as experimental methodology to study transplant and tumor biology. Nat Protoc 4, 86–93 (2009). https://doi.org/10.1038/nprot.2008.218

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