Development of human alveolar epithelial cell models to study distal lung biology and disease

Evelyn Tran; Tuo Shi; Xiuwen Li; Adnan Y Chowdhury; Du Jiang; Yixin Liu; Hongjun Wang; Chunli Yan; William D Wallace; Rong Lu; Amy L Ryan; Crystal N Marconett; Beiyun Zhou; Zea Borok; Ite A Offringa

doi:10.1016/j.isci.2022.103780

Development of human alveolar epithelial cell models to study distal lung biology and disease

iScience. 2022 Jan 15;25(2):103780. doi: 10.1016/j.isci.2022.103780. eCollection 2022 Feb 18.

Authors

Evelyn Tran^{1

2

3}, Tuo Shi^{1

2

3}, Xiuwen Li^{2

4}, Adnan Y Chowdhury⁵, Du Jiang⁵, Yixin Liu⁶, Hongjun Wang⁶, Chunli Yan^{1

2}, William D Wallace⁷, Rong Lu⁵, Amy L Ryan^{5

6}, Crystal N Marconett^{1

2

3}, Beiyun Zhou^{2

6}, Zea Borok^{2

3

6

8}, Ite A Offringa^{1

2

3}

Affiliations

¹ Department of Surgery, Keck School of Medicine, University of Southern California (USC), Los Angeles, CA 90033, USA.
² USC Norris Comprehensive Cancer Center, Keck School of Medicine, USC, Los Angeles, CA 90033, USA.
³ Department of Biochemistry and Molecular Medicine, Keck School of Medicine, USC, Los Angeles, CA 90033, USA.
⁴ Department of Translational Genomics, Keck School of Medicine, USC, Los Angeles, CA 90033, USA.
⁵ Department of Stem Cell Biology and Regenerative Medicine, Eli and Edythe Broad CIRM Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, USC, Los Angeles, CA 90033, USA.
⁶ Hastings Center for Pulmonary Research and Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Keck School of Medicine, USC, Los Angeles, CA 90033, USA.
⁷ Department of Pathology, Keck School of Medicine, USC, Los Angeles, CA 90033, USA.
⁸ Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, University of California, San Diego, La Jolla, CA 92093, USA.

Abstract

Many acute and chronic diseases affect the distal lung alveoli. Alveolar epithelial cell (AEC) lines are needed to better model these diseases. We used de-identified human remnant transplant lungs to develop a method to establish AEC lines. The lines grow well in 2-dimensional (2D) culture as epithelial monolayers expressing lung progenitor markers. In 3-dimensional (3D) culture with fibroblasts, Matrigel, and specific media conditions, the cells form alveolar-like organoids expressing mature AEC markers including aquaporin 5 (AQP5), G-protein-coupled receptor class C group 5 member A (GPRC5A), and surface marker HTII280. Single-cell RNA sequencing of an AEC line in 2D versus 3D culture revealed increased cellular heterogeneity and induction of cytokine and lipoprotein signaling in 3D organoids. Our approach yields lung progenitor lines that retain the ability to differentiate along the alveolar cell lineage despite long-term expansion and provides a valuable system to model and study the distal lung in vitro.

Keywords: Cell biology; Developmental biology; Stem cells research; Transcriptomics.

Abstract

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