Acellular Normal and Fibrotic Human Lung Matrices as a Culture System for In Vitro Investigation
Open accessAbstract
Rationale: Extracellular matrix (ECM) is a dynamic tissue that contributes to organ integrity and function, and its regulation of cell phenotype is a major aspect of cell biology. However, standard in vitro culture approaches are of unclear physiologic relevance because they do not mimic the compositional, architectural, or distensible nature of a living organ. In the lung, fibroblasts exist in ECM-rich interstitial spaces and are key effectors of lung fibrogenesis. Objectives: To better address how ECM influences fibroblast phenotype in a disease-specific manner, we developed a culture system using acellular human normal and fibrotic lungs. Methods: Decellularization was achieved using treatment with detergents, salts, and DNase. The resultant matrices can be sectioned as uniform slices within which cells were cultured. Measurements and Main Results: We report that the decellularization process effectively removes cellular and nuclear material while retaining native dimensionality and stiffness of lung tissue. We demonstrate that lung fibroblasts reseeded into acellular lung matrices can be subsequently assayed using conventional protocols; in this manner we show that fibrotic matrices clearly promote transforming growth factor-β–independent myofibroblast differentiation compared with normal matrices. Furthermore, comprehensive analysis of acellular matrix ECM details significant compositional differences between normal and fibrotic lungs, paving the way for further study of novel hypotheses. Conclusions: This methodology is expected to allow investigation of important ECM-based hypotheses in human tissues and permits future scientific exploration in an organ- and disease-specific manner.
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Alterations in emphysematous or fibrotic lung tissue can lead to cells failing to survive beyond one week of culture or broad changes in the phenotype of seeded cells that do survive.
“These alterations can lead to cells failing to survive beyond one week of culture118 or broad changes in the phenotype of seeded cells that do survive119.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [119]
Emphysematous or fibrotic lung tissue has hardened and undergone alterations in its architecture.
“For example, emphysematous or fibrotic lung tissue has hardened and undergone alterations in its architecture.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [119]
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Unravelling the memory of the extracellular matrix using MASH-derived decellularized scaffolds
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“…cues or “zip codes” direct the behavior of resident and interacting cells, such that homeostatic matrix properties promote normal tissue development and function [13, 14], whereas altered or diseased matrix states drive maladaptive cellular responses and diseased tissue formation [10, 12, 15].”
- Authors
- Booth, A. J., Hadley, R., Cornett, A. M., Dreffs, A. A., Matthes, S. A., Tsui, J. L., Weiss, K., Horowitz, J. C., Fiore, V. F., Barker, T. H., Moore, B. B., Martinez, F. J., Niklason, L. E., & White, E. S.
- Affiliations
- University of Michigan (US)
- Georgia Institute of Technology (US)
- Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan
- Yale University (US)
- Corresponding author
- White, Eric S.
- ORCID
- Venue
- American Journal of Respiratory and Critical Care Medicine
- Publisher
- Oxford University Press (OUP)
- Published
- 2012-11-01
- Volume
- 186
- Issue
- 9
- Pages
- 866-876
- ISSN
- 1073-449X (ISSN-L)
- 1535-4970
- PMC
- PMC3530219
- OpenAlex
- W2062461864
- Cited by
- 665
- References
- 53
- Citations by year
- 2026: 36 · 2025: 50 · 2024: 46 · 2023: 63 · 2022: 57 · 2021: 64 · 2020: 49 · 2019: 37 · 2018: 59 · 2017: 53 · 2016: 44 · 2015: 49 · 2014: 33 · 2013: 22 · 2012: 3
- FWCI
- 34.94
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- American Journal of Respiratory and Critical Care Medicine (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Health Effects Institute
- Georgia Institute of Technology
- National Institutes of Health
- National Heart, Lung, and Blood Institute (R01HL098220, T32HL07749, U01HL111016, R01HL085083, R01HL105489, T32HL007749)
- Topics
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Interstitial Lung Diseases and Idiopathic Pulmonary Fibrosis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Keywords
- decellularization, pulmonary fibrosis, lung fibroblasts, extracellular matrix, myofibroblast differentiation, lung stiffness, matrix composition, in vitro culture system, organotypic culture
- MeSH terms
- Extracellular Matrix, Fibroblasts, Humans, Lung, Microscopy, Electron, Pulmonary Fibrosis, Spectrophotometry, Atomic, Mass Spectrometry, Blotting, Western, Tissue Culture Techniques
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