title: Differentiation of Club Cells to Alveolar Epithelial Cells In Vitro.
abstract: Club cells are known to function as regional progenitor cells to repair the bronchiolar epithelium in response to lung damage. By lineage tracing in mice, we have shown recently that club cells also give rise to alveolar type 2 cells (AT2s) and alveolar type 1 cells (AT1s) during the repair of the damaged alveolar epithelium. Here, we show that when highly purified, anatomically and phenotypically confirmed club cells are seeded in 3-dimensional culture either in bulk or individually, they proliferate and differentiate into both AT2- and AT1-like cells and form alveolar-like structures. This differentiation was further confirmed by transcriptomic analysis of freshly isolated club cells and their cultured progeny. Freshly isolated club cells express Sca-1 and integrin α6, markers commonly used to characterize lung stem/progenitor cells. Together, current study for the first time isolated highly purified club cells for in vitro study and demonstrated club cells' capacity to differentiate into alveolar epithelial cells at the single-cell level.
authors.affiliation: Interdisciplinary Research Group in Infectious Diseases, Singapore-Massachusetts Institute of Technology Alliance for Research and Technology, Singapore.|A*STAR Institute of Molecular and Cell Biology, 61 Biopolis Drive Proteos, Singapore 138673, Singapore.
authors.forename: Dahai
authors.initials: D
authors.lastname: Zheng
authors.affiliation: A*STAR Institute of Molecular and Cell Biology, 61 Biopolis Drive Proteos, Singapore 138673, Singapore.|Department of Biological Sciences, National University of Singapore, 117543, Singapore.
authors.forename: Boon-Seng
authors.initials: BS
authors.lastname: Soh
authors.affiliation: Interdisciplinary Research Group in BioSystems and Micromechanics, Singapore-Massachusetts Institute of Technology Alliance for Research and Technology, Singapore.
authors.forename: Lu
authors.initials: L
authors.lastname: Yin
authors.affiliation: The Koch Institute for Integrative Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Guangan
authors.initials: G
authors.lastname: Hu
authors.affiliation: A*STAR Institute of Molecular and Cell Biology, 61 Biopolis Drive Proteos, Singapore 138673, Singapore.
authors.forename: Qingfeng
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Saw Swee Hock School of Public Health, National University of Singapore, Singapore.
authors.forename: Hyungwon
authors.initials: H
authors.lastname: Choi
authors.affiliation: Interdisciplinary Research Group in BioSystems and Micromechanics, Singapore-Massachusetts Institute of Technology Alliance for Research and Technology, Singapore.|Department of Electrical Engineering and Computer Science, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Jongyoon
authors.initials: J
authors.lastname: Han
authors.affiliation: Host and Pathogen Interactivity Laboratory, Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Health System, National University of Singapore, Singapore.
authors.forename: Vincent T K
authors.initials: VT
authors.lastname: Chow
authors.affiliation: Interdisciplinary Research Group in Infectious Diseases, Singapore-Massachusetts Institute of Technology Alliance for Research and Technology, Singapore.|The Koch Institute for Integrative Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Jianzhu
authors.initials: J
authors.lastname: Chen
mesh_terms: D056809:Alveolar Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D000818:Animals; D000067696:Ataxin-1 / Q000378:metabolism; D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016130:Immunophenotyping; D039503:Integrin alpha6 / Q000378:metabolism; D051379:Mice; D010641:Phenotype; D020545:Respiratory Mucosa / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D059467:Transcriptome
chemical_list: D000067696:Ataxin-1; D015415:Biomarkers; D039503:Integrin alpha6; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins