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Preparation of Decellularized Lung Matrices for Cell Culture and Protein Analysis

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Fibrosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1627))

Abstract

The limited available treatment options for patients with chronic lung diseases, such as fibrosis, lead to poor prognosis after diagnosis and short survival rates. An exciting new bioengineering approach utilizes de- and recellularization of lung tissue to potentially overcome donor organ shortage and immune reactions toward the received transplant. The goal of decellularization is to create a scaffold which contains the necessary framework for stability and functionality for regenerating lung tissue while removing immunomodulatory factors by removal of cells. After decellularization, the scaffold could be re-functionalized by repopulation with the patient’s own stem/progenitor cells to create a fully functional organ or can be used as ex vivo models of disease. In this chapter the decellularization of lung tissue from multiple species (i.e., rodents, pigs, and humans) as well as disease states such as fibrosis is described. We discuss and describe the various quality control measures which should be used to characterize decellularized scaffolds, methods for protein analysis of the remaining scaffold, and methods for recellularization of scaffolds.

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Acknowledgments

The authors are grateful to Douglas Taatjes Ph.D. and Nicole Bishop of the UVM Microscopy and Imaging Center for assistance with IHC staining, transmission electron microscopy imaging, and image interpretations. We would like to thank Caroline Andrews, Nicholas Bonenfant, Zachary Borg, Amy Coffey, Amanda Daly, Jacob Dearborn, Nathan Gasek, Ethan Griswold, Zachary Phillips, Joseph Platz, Alexander Riveron, Dino Sokocevic, John Wallis, Sean Wrenn, and Basa Zvorova for the assistance with lung decellularizations, microscopy, and analysis of the detergent effluents. Studies were supported by the NIH RO1 HL127144-01 (DJW), Vermont Lung Center CoBRE grant (P20RR15557), NIH PACT program (contract HHSN268201000008C), NHBLI Lung Biology Training grant T32 HL076122, NIH Institutional Development Award (IDeA) NIGMS grant P20GM103449, and NCRR grant P40RR017447.

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Authors and Affiliations

  1. Department of Med-Pulmonary, College of Medicine, University of Vermont, Burlington, VT, USA

    Franziska E. Uhl & Daniel J. Weiss

  2. Comprehensive Pneumology Center, University Hospital, Ludwig-Maximilians University, Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Munich, Germany

    Darcy E. Wagner

Authors
  1. Franziska E. Uhl
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  2. Darcy E. Wagner
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  3. Daniel J. Weiss
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Corresponding author

Correspondence to Franziska E. Uhl .

Editor information

Editors and Affiliations

  1. Department of Dermatology, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Laure Rittié

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Uhl, F.E., Wagner, D.E., Weiss, D.J. (2017). Preparation of Decellularized Lung Matrices for Cell Culture and Protein Analysis. In: Rittié, L. (eds) Fibrosis. Methods in Molecular Biology, vol 1627. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7113-8_18

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  • DOI: https://doi.org/10.1007/978-1-4939-7113-8_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7112-1

  • Online ISBN: 978-1-4939-7113-8

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