In vitro expansion of endogenous human alveolar epithelial type II cells in fibroblast-free spheroid culture

Biochem Biophys Res Commun. 2019 Aug 6;515(4):579-585. doi: 10.1016/j.bbrc.2019.05.187. Epub 2019 Jun 6.

Abstract

Alveolar epithelial type II cells (AEC2) are stem cells of the alveoli and play crucial roles in maintaining lung homeostasis and the pathogenesis of lung diseases. We recently reported on an organoid culture system for endogenous murine AEC2. Despite advances in generation of human induced pluripotent stem cell-derived AEC2, in vitro expansion of endogenous human AEC2 has not been reported and genetic manipulation of human AEC2 has been difficult. Here, we show that endogenous human AEC2 could be cultured and passaged using a three-dimensional culture system with a specific combination of signal ligands and inhibitors. The culture system was suitable for retroviral gene transduction into AEC2. Transduction of pulmonary fibrosis-associated mutant surfactant protein C (SFPTCΔexon4) into AEC2 revealed characteristic transcriptional traits similar to those of patients with idiopathic pulmonary fibrosis. Our culture system will be a useful tool for investigating human AEC2 functions in vitro.

Keywords: Alveolar epithelial cell; Fibroblast; Flow cytometry; Lung; Primary cell culture; Transcriptome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alveolar Epithelial Cells / cytology*
  • Cell Culture Techniques*
  • Cell Line
  • Cells, Cultured
  • Fibroblasts
  • Fibrosis
  • Gene Expression Profiling
  • Humans
  • Hyperoxia / metabolism
  • Induced Pluripotent Stem Cells / cytology
  • Ligands
  • Male
  • Mutation
  • Pulmonary Alveoli / cytology*
  • Pulmonary Surfactant-Associated Protein C / genetics*
  • Spheroids, Cellular / cytology*
  • Transcription, Genetic
  • Transcriptome

Substances

  • Ligands
  • Pulmonary Surfactant-Associated Protein C
  • SFTPC protein, human