A medium composition containing normal resting glucose that supports differentiation of primary human airway cells

• Type of Material: Article
• Authors: Rachel Morgan, Emory University; Candela Manfredi, Emory University; Kristen F. Easley, Emory University; Lionel D. Watkins, Emory University; William Hunt, Emory University; Steven Goudy, Emory University; Eric Sorscher, Emory University; Michael Koval, Emory University; Samuel Molina, Emory University
• Language: English
• Date: 2022-01-27
• Publisher: NATURE PORTFOLIO
• Publication Version: Final Published Version
• Copyright Statement: © The Author(s) 2022
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/s41598-022-05446-x
• Title of Journal or Parent Work: SCIENTIFIC REPORTS
• Volume: 12
• Issue: 1
• Start Page: 1540
• End Page: 1540
• Grant/Funding Information: This research project was supported by the Emory University Integrated Cellular Imaging Microscopy Core, the Winship Research Pathology Core and SEM was done by the Robert P. Apkarian Integrated Electron Microscopy Core (IEMC) of Emory University School of Medicine. This project was also supported by CF@LANTA RDP Center Grant (MCCART15R0), R01-HL116958 and R01-AA025854 to M.K., T32-HL116271 to S.A.M., R01-HL139876 and R01-HL136414 to E.J.S., F31-AA029000 to K.F.E., and by the Emory+Children’s Center for Cystic Fibrosis and Airways Disease Research.
• Keywords: MODEL; CYSTIC-FIBROSIS PATIENT; Multidisciplinary Sciences; LONG-TERM EXPANSION; Science & Technology; INHIBITOR; IN-VITRO; METABOLISM; VIVO; Science & Technology - Other Topics; EPITHELIAL-CELLS
• Research Categories: Health Sciences, Nutrition; Health Sciences, Obstetrics and Gynecology; Biology, Cell

Abstract

Primary cells isolated from the human respiratory tract are the state-of-the-art for in vitro airway epithelial cell research. Airway cell isolates require media that support expansion of cells in a basal state to maintain the capacity for differentiation as well as proper cellular function. By contrast, airway cell differentiation at an air–liquid interface (ALI) requires a distinct medium formulation that typically contains high levels of glucose. Here, we expanded and differentiated human basal cells isolated from the nasal and conducting airway to a mature mucociliary epithelial cell layer at ALI using a medium formulation containing normal resting glucose levels. Of note, bronchial epithelial cells expanded and differentiated in normal resting glucose medium showed insulin-stimulated glucose uptake which was inhibited by high glucose concentrations. Normal glucose containing ALI also enabled differentiation of nasal and tracheal cells that showed comparable electrophysiological profiles when assessed for cystic fibrosis transmembrane conductance regulator (CFTR) function and that remained responsive for up to 7 weeks in culture. These data demonstrate that normal glucose containing medium supports differentiation of primary nasal and lung epithelial cells at ALI, is well suited for metabolic studies, and avoids pitfalls associated with exposure to high glucose.