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Bioengineering

Dissection and Isolation of Region-Specific Decellularized Lung Tissue

Published: September 29th, 2023

DOI:

10.3791/65276

1Larner College of Medicine, University of Vermont, 2College of Engineering and Mathematical Sciences, University of Vermont

Presented here is a protocol for the isolation of regional decellularized lung tissue. This protocol provides a powerful tool for studying complexities in the extracellular matrix and cell-matrix interactions.

Lung transplantation is often the only option for patients in the later stages of severe lung disease, but this is limited both due to the supply of suitable donor lungs and both acute and chronic rejection after transplantation. Ascertaining novel bioengineering approaches for the replacement of diseased lungs is imperative for improving patient survival and avoiding complications associated with current transplantation methodologies. An alternative approach involves the use of decellularized whole lungs lacking cellular constituents that are typically the cause of acute and chronic rejection. Since the lung is such a complex organ, it is of interest to examine the extracellular matrix components of specific regions, including the vasculature, airways, and alveolar tissue. The purpose of this approach is to establish simple and reproducible methods by which researchers may dissect and isolate region-specific tissue from fully decellularized lungs. The current protocol has been devised for pig and human lungs, but may be applied to other species as well. For this protocol, four regions of the tissue were specified: airway, vasculature, alveoli, and bulk lung tissue. This procedure allows for the procurement of samples of tissue that more accurately represent the contents of the decellularized lung tissue as opposed to traditional bulk analysis methods.

Lung diseases, including chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), and cystic fibrosis (CF), currently remain without a cure1,2,3,4. Lung transplantation is often the only option for patients in later stages, however this remains a limited option both due to the supply of suitable donor lungs and both acute and chronic rejection after transplantation3,5,6. As such, there is a critical need for new treatment s....

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All animal studies have been performed in accordance with the IACUC of University of Vermont (UVM). All human lungs were acquired from UVM Autopsy Services and related studies were performed as per the guidelines of IRB of UVM.

NOTE: Decellularization of pig and human lungs has been previously described by our group7,8,9,10, 21. In .......

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An overall schematic of the protocol is depicted in Figure 1. Once mastered, the regional dissection of decellularized lung tissue is easily reproducible. Determining the categorization of each severed tissue sample is imperative to the success of the dissection procedure. Vascular tissue is substantially more elastic than airway, so using forceps to stretch the tissue is often a strong indicator of whether a particular sample is vasculature or airway. Typically, vascular tissue runs paralle.......

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Decellularized tissues from humans and other species are frequently utilized as biomaterials for studying ECM composition as well as cell-ECM interactions in ex vivo culture models, including 3D hydrogels12,13. Similar to other organs, decellularized lungs have previously been utilized to determine ECM compositional differences in healthy versus diseased (i.e., emphysematous and IPF) lungs and are increasingly being utilized as hydrogels for studying ECM.......

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The authors thank the UVM autopsy services for human lung procurement and Robert Pouliot PhD for contributions to the overall dissection techniques. These studies were supported by R01 HL127144-01 (DJW).

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Name Company Catalog Number Comments
Bonn Scissors Fine Science Tools 14184-09
Dumont #5 - Fine Forceps Fine Science Tools 11254-02
Forceps, Curved, S/S, Blunt, Serrated - 130mm CellPath N/A
Hardened Fine Scissors Fine Science Tools 14090-11
Moria Iris Forceps Fine Science Tools 11373-22
Pyrex Glass Casserole Dish Cole-Parmer 3175-10

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