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Isolation of epithelial, endothelial, and immune cells from lungs of transgenic mice with oncogene-induced lung adenocarcinomas.

Abstract
Genetically engineered mouse models of lung adenocarcinoma have proven invaluable for understanding mechanisms of tumorigenesis, therapy response, and drug resistance. However, mechanistic studies focused on studying these processes in tumor-bearing mouse lungs are confounded by the fact that, in most cases, relevant signaling pathways are analyzed in whole-lung preparations, which are composed of a heterogeneous mixture of cells. Given our increasing knowledge about the roles played by different subpopulations of cells in the development of lung adenocarcinoma, separating the major cellular compartments of the tumor microenvironment is recommended to allow for a precise analysis of relevant pathways in each isolated cell type. In this study, we optimized magnetic- and fluorescence-based isolation protocols to segregate lung epithelial (CD326/epithelial cell adhesion molecule-positive), endothelial (CD31-positive), and immune (CD45-positive) cells, with high purity, from the lungs of transgenic mice with mutant epidermal growth factor receptor-induced lung adenocarcinomas. This approach, which can potentially be extended to additional lung adenocarcinoma models, enables delineation of the molecular features of individual cell types that can be used to gain insight into their roles in lung adenocarcinoma initiation, progression, and response to therapy.
AuthorsAmlak Bantikassegn, Xiaoling Song, Katerina Politi
JournalAmerican journal of respiratory cell and molecular biology (Am J Respir Cell Mol Biol) Vol. 52 Issue 4 Pg. 409-17 (Apr 2015) ISSN: 1535-4989 [Electronic] United States
PMID25347711 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
Chemical References
  • EGFR protein, mouse
  • ErbB Receptors
Topics
  • Adenocarcinoma (genetics, immunology, pathology)
  • Adenocarcinoma of Lung
  • Animals
  • Cells, Cultured
  • Disease Models, Animal
  • Endothelial Cells (physiology)
  • Epithelial Cells (physiology)
  • ErbB Receptors (genetics)
  • Flow Cytometry
  • Humans
  • Immunomagnetic Separation
  • Lung (immunology, pathology)
  • Lung Neoplasms (genetics, immunology, pathology)
  • Macrophages (physiology)
  • Mice, Transgenic
  • Oncogenes
  • Respiratory Mucosa (pathology)

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