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The maintenance of lung function relies crucially on the homeostatic replacement and post-injury regeneration of the lung alveolar epithelium (Hogan et al., Cell Stem Cell 15(2):123–138, 2014). Dysfunctions in these processes contribute to the pathogenesis of numerous lung diseases (Hogan et al., Cell Stem Cell 15(2):123–138, 2014; Matthay et al., J Clin Invest 122(8):2731–2740, 2012). While identifying stem and progenitor cells in the lung epithelium has significantly enriched our understanding of endogenous replacement and regenerative mechanisms (Hogan et al., Cell Stem Cell 15(2):123–138, 2014), it is clear that epithelial cells interact closely with mesenchymal components which create a micro-environmental niche that is vital for regulating both homeostatic replacement and post-injury regeneration of epithelial cells. Specific subsets of alveolar type II cells (AT2) behave as epithelial stem cells of the distal lung. We have identified a CD44high subpopulation of AT2 cells that are preferentially located near macro-blood vessels and manifest stem cell characteristics (Chen et al., Stem Cell Rep 19(6):890–905, 2024; Am J Physiol Lung Cell Mol Physiol 313(1):L41–l51, 2017). In addition, the macro-blood vessels endothelial cells (ECs) function as niche components to support the CD44high AT2s. Here, we describe the method to identify the CD44high AT2 cells by immuno-fluorescence and co-culture of CD44high AT2 cells with lung endothelial cells—their potential niche component—in 3D organoid culture.

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Co-colonization and Co-culture of Lung Alveoli Epithelial Stem Cells and Their Endothelial Niche Cells

  • 实验方案

  • 发表日期：

    2025年

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  • Hiroyuki Hirai,
  • Qian Chen,
  • Yuru Liu

摘要

The maintenance of lung function relies crucially on the homeostatic replacement and post-injury regeneration of the lung alveolar epithelium (Hogan et al., Cell Stem Cell 15(2):123–138, 2014). Dysfunctions in these processes contribute to the pathogenesis of numerous lung diseases (Hogan et al., Cell Stem Cell 15(2):123–138, 2014; Matthay et al., J Clin Invest 122(8):2731–2740, 2012). While identifying stem and progenitor cells in the lung epithelium has significantly enriched our understanding of endogenous replacement and regenerative mechanisms (Hogan et al., Cell Stem Cell 15(2):123–138, 2014), it is clear that epithelial cells interact closely with mesenchymal components which create a micro-environmental niche that is vital for regulating both homeostatic replacement and post-injury regeneration of epithelial cells. Specific subsets of alveolar type II cells (AT2) behave as epithelial stem cells of the distal lung. We have identified a CD44high subpopulation of AT2 cells that are preferentially located near macro-blood vessels and manifest stem cell characteristics (Chen et al., Stem Cell Rep 19(6):890–905, 2024; Am J Physiol Lung Cell Mol Physiol 313(1):L41–l51, 2017). In addition, the macro-blood vessels endothelial cells (ECs) function as niche components to support the CD44high AT2s. Here, we describe the method to identify the CD44high AT2 cells by immuno-fluorescence and co-culture of CD44high AT2 cells with lung endothelial cells—their potential niche component—in 3D organoid culture.