Preparation of Decellularized Lung Matrices for Cell Culture and Protein Analysis

Methods Mol Biol. 2017:1627:253-283. doi: 10.1007/978-1-4939-7113-8_18.

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.

Keywords: Biomaterial; Decellularization; Lung; Mass spectrometry; Proteomics; Scaffold; Tissue engineering.

MeSH terms

  • Animals
  • Bleomycin / adverse effects
  • Bleomycin / analogs & derivatives
  • Cell Culture Techniques*
  • Disease Models, Animal
  • Endothelial Cells / metabolism
  • Extracellular Matrix
  • Fibroblasts / metabolism
  • Humans
  • Immunohistochemistry
  • Lung / metabolism
  • Lung / pathology
  • Lung / ultrastructure
  • Mass Spectrometry
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Proteomics* / methods
  • Pulmonary Fibrosis / chemically induced
  • Pulmonary Fibrosis / metabolism
  • Pulmonary Fibrosis / pathology
  • Regeneration
  • Swine
  • Tissue Engineering* / methods
  • Tissue Scaffolds*

Substances

  • Bleomycin
  • bleomycetin