A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface
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...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2014-01-01
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| Series: | Redox Biology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231714000196 |
| _version_ | 1828417660777398272 |
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| author | Weiling Xu Allison J. Janocha Rachel A. Leahy Ryan Klatte Dave Dudzinski Lori A. Mavrakis Suzy A.A. Comhair Mark E. Lauer Calvin U. Cotton Serpil C. Erzurum |
| author_facet | Weiling Xu Allison J. Janocha Rachel A. Leahy Ryan Klatte Dave Dudzinski Lori A. Mavrakis Suzy A.A. Comhair Mark E. Lauer Calvin U. Cotton Serpil C. Erzurum |
| author_sort | Weiling Xu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T14:23:27Z |
| format | Article |
| id | doaj.art-8345b9e109ab41ce92c9982bea3125a6 |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-12-10T14:23:27Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-8345b9e109ab41ce92c9982bea3125a62022-12-22T01:45:08ZengElsevierRedox Biology2213-23172014-01-012C51351910.1016/j.redox.2014.01.004A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interfaceWeiling Xu0Allison J. Janocha1Rachel A. Leahy2Ryan Klatte3Dave Dudzinski4Lori A. Mavrakis5Suzy A.A. Comhair6Mark E. Lauer7Calvin U. Cotton8Serpil C. Erzurum9Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USADivision of Pediatric Pulmonology, Case Western University, Cleveland, OH 44106, USALerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USAAir–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.http://www.sciencedirect.com/science/article/pii/S2213231714000196Airway epithelial cellsAir–liquid interface cultureMetabolismOxygen consumption rate (OCR)Extracellular acidification rate (ECAR) |
| spellingShingle | Weiling Xu Allison J. Janocha Rachel A. Leahy Ryan Klatte Dave Dudzinski Lori A. Mavrakis Suzy A.A. Comhair Mark E. Lauer Calvin U. Cotton Serpil C. Erzurum A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface Redox Biology Airway epithelial cells Air–liquid interface culture Metabolism Oxygen consumption rate (OCR) Extracellular acidification rate (ECAR) |
| title | A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface |
| title_full | A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface |
| title_fullStr | A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface |
| title_full_unstemmed | A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface |
| title_short | A novel method for pulmonary research: Assessment of bioenergetic function at the air–liquid interface |
| title_sort | novel method for pulmonary research assessment of bioenergetic function at the air liquid interface |
| topic | Airway epithelial cells Air–liquid interface culture Metabolism Oxygen consumption rate (OCR) Extracellular acidification rate (ECAR) |
| url | http://www.sciencedirect.com/science/article/pii/S2213231714000196 |
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