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

• Published in: Scientific reports Vol. 12; no. 1; pp. 1540 - 15
• Main Authors: Morgan, Rachel, Manfredi, Candela, Easley, Kristen F., Watkins, Lionel D., Hunt, William R., Goudy, Steven L., Sorscher, Eric J., Koval, Michael, Molina, Samuel A.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 27.01.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/1407/651; 631/80; 631/80/304; 692/699/1785/4039; Basal cells; Bronchi - cytology; Bronchi - metabolism; Cell culture; Cell Culture Techniques - methods; Cell differentiation; Cell Differentiation - drug effects; Cells, Cultured; Culture Media; Cystic fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator - metabolism; Epithelial cells; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Glucose; Glucose - metabolism; Glucose - pharmacology; Humanities and Social Sciences; Humans; Insulin; multidisciplinary; Respiratory Mucosa - cytology; Respiratory Mucosa - drug effects; Respiratory Mucosa - metabolism; Respiratory tract; Science; Science (multidisciplinary); Trachea - cytology; Trachea - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-05446-x

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.