title: A novel method for pulmonary research: assessment of bioenergetic function at the air-liquid interface.
abstract: Air-liquid interface cell culture is an organotypic model for study of differentiated functional airway epithelium in vitro. Dysregulation of cellular energy metabolism and mitochondrial function have been suggested to contribute to airway diseases. However, there is currently no established method to determine oxygen consumption and glycolysis in airway epithelium in air-liquid interface. In order to study metabolism in differentiated airway epithelial cells, we engineered an insert for the Seahorse XF24 Analyzer that enabled the measure of respiration by oxygen consumption rate (OCR) and glycolysis by extracellular acidification rate (ECAR). Oxidative metabolism and glycolysis in airway epithelial cells cultured on the inserts were successfully measured. The inserts did not affect the measures of OCR or ECAR. Cells under media with apical and basolateral feeding had less oxidative metabolism as compared to cells on the inserts at air-interface with basolateral feeding. The design of inserts that can be used in the measure of bioenergetics in small numbers of cells in an organotypic state may be useful for evaluation of new drugs and metabolic mechanisms that underlie airway diseases.
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Weiling
authors.initials: W
authors.lastname: Xu
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Allison J
authors.initials: AJ
authors.lastname: Janocha
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Leahy
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Klatte
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Dave
authors.initials: D
authors.lastname: Dudzinski
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Lori A
authors.initials: LA
authors.lastname: Mavrakis
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Suzy A A
authors.initials: SA
authors.lastname: Comhair
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Lauer
authors.affiliation: Division of Pediatric Pulmonology, Case Western University, Cleveland, OH 44106, USA.
authors.forename: Calvin U
authors.initials: CU
authors.lastname: Cotton
authors.affiliation: Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA ; Respiratory Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Serpil C
authors.initials: SC
authors.lastname: Erzurum
mesh_terms: D000328:Adult; D004734:Energy Metabolism; D005080:Exercise Test / Q000379:methods*; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D007717:Knee / Q000502:physiology*; D008168:Lung / Q000502:physiology; D008297:Male; D018482:Muscle, Skeletal / Q000502:physiology*; D018384:Oxidative Stress; D005082:Physical Exertion; D017382:Reactive Oxygen Species / Q000097:blood / Q000652:urine; D055815:Young Adult
chemical_list: D017382:Reactive Oxygen Species; D005978:Glutathione
keywords: Airway epithelial cells; Air–liquid interface culture; ECAR, extracellular acidification rate; Extracellular acidification rate (ECAR); MTEC, mouse tracheal epithelial cell; Metabolism; OCR, oxygen consumption rate; Oxygen consumption rate (OCR); TCA, tricarboxylic acid; iNOS, inducible nitric oxide synthase