A novel method for isolating individual cellular components from the adult human distal lung

Naoya Fujino, Hiroshi Kubo, Chiharu Ota, Takaya Suzuki, Satoshi Suzuki, Mitsuhiro Yamada, Toru Takahashi, Mei He, Takashi Suzuki, Takashi Kondo, Mutsuo Yamaya

Research output: Contribution to journalArticlepeer-review

46 Citations (Scopus)

Abstract

A variety of lung diseases, such as pulmonary emphysema and idiopathic pulmonary fibrosis, develop in the lung alveoli. Multiple cell types are localized in the alveoli, including epithelial, mesenchymal, and endothelial cells. These resident cells participate in the pathogenesis of lung disease in various ways. To elaborate clearly on the mechanisms of these pathologic processes, cell type-specific analyses of lung disease are required. However, no method exists for individually isolating the different types of cells found in the alveoli. We report on the development of a FACS-based method for the direct isolation of individual cell types from the adult human distal lung.Weobtained human lung tissue from lung resections, and prepared single-cell suspension. After depleting CD45-positive cells, a combination of antibodies against epithelial cell adhesion molecule( EpCAM),T1α, and vascular endothelial (VE)-cadherin as used to delineate alveolar cell types. Alveolar Type II cells were highly purified in the EpCAM hi/T1α - subset, whereas the EpCAM +/T1α -/low subset contained a mixed epithelial population consisting of alveolar Type I and bronchiolar epithelial cells. The EpCAM -/T1α - subset included both microvascular endothelial and mesenchymal cells, and these were separated by immunoreactivity to VE-cadherin. Lymphatic endothelial cells existed in the EpCAM -/T1α hi subset. Isolated cells were viable, and further cell culture studies could be performed. These results suggest that this novel method enables the isolation of different cellular components from normal and diseased lungs, and is capable of elucidating phenotypes specific to certain alveolar cell types indicative of lung disease.

Original languageEnglish
Pages (from-to)422-430
Number of pages9
JournalAmerican journal of respiratory cell and molecular biology
Volume46
Issue number4
DOIs
Publication statusPublished - 2012 Apr

Keywords

  • Cell culture
  • Cell isolation
  • Fluorescence-activated cell sorting
  • Pulmonary alveoli

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

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