Back to Search Start Over

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

Authors :
Booth AJ
Hadley R
Cornett AM
Dreffs AA
Matthes SA
Tsui JL
Weiss K
Horowitz JC
Fiore VF
Barker TH
Moore BB
Martinez FJ
Niklason LE
White ES
Source :
American journal of respiratory and critical care medicine [Am J Respir Crit Care Med] 2012 Nov 01; Vol. 186 (9), pp. 866-76. Date of Electronic Publication: 2012 Aug 30.
Publication Year :
2012

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.<br />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.<br />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.<br />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.<br />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.

Details

Language :
English
ISSN :
1535-4970
Volume :
186
Issue :
9
Database :
MEDLINE
Journal :
American journal of respiratory and critical care medicine
Publication Type :
Academic Journal
Accession number :
22936357
Full Text :
https://doi.org/10.1164/rccm.201204-0754OC