Acellular normal and fibrotic human lung matrices as a culture system for in vitro investigation

Adam J. Booth, Ryan Hadley, Ashley M. Cornett, Alyssa A. Dreffs, Stephanie A. Matthes, Jessica L. Tsui, Kevin Weiss, Jeffrey C. Horowitz, Vincent F. Fiore, Thomas H. Barker, Bethany B. Moore, Fernando J. Martinez, Laura E. Niklason, Eric S. White*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

334 Scopus citations

Abstract

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.Wedemonstrate that lung fibroblasts reseeded into acellular lung matrices can be subsequently assayed using conventional protocols; in this mannerweshowthat fibrotic matrices clearlypromotetransforming growth factor-b-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.

Original languageEnglish (US)
Pages (from-to)866-876
Number of pages11
JournalAmerican journal of respiratory and critical care medicine
Volume186
Issue number9
DOIs
StatePublished - Nov 1 2012

Keywords

  • Extracellular matrix
  • Fibroblast
  • Lung fibrosis

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Critical Care and Intensive Care Medicine

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