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 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.

• Publication year

  2014

• Venue

  Redox Biology

• Publication date

  2014-01-16

• Fields of study

  Biology, Medicine, Environmental Science

• Identifiers
  DOI 10.1016/j.redox.2014.01.004PMID 24624341PMCID 3949089
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• The insert design supports bioenergetics measurements in small numbers of cells in an organotypic state and may be useful for evaluating drugs and metabolic mechanisms relevant to airway diseases.

  Confidence 0.88

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• Cells under apical and basolateral feeding showed lower oxidative metabolism than cells on inserts at the air-interface with basolateral feeding.

  Confidence 0.91

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• The inserts did not alter OCR or ECAR measurements.

  Confidence 0.95

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• An engineered insert for the Seahorse XF24 Analyzer enabled measurement of respiration via oxygen consumption rate (OCR) and glycolysis via extracellular acidification rate (ECAR) in differentiated airway epithelial cells at an air-liquid interface.

  Confidence 0.98

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review

• air-interface with basolateral feeding

  culture condition

  A culture condition in which airway epithelial cells are maintained at an air interface with medium supplied basolaterally.

  Aliases: air-interface basolateral feeding

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• air-liquid interface cell culture

  culture system

  An organotypic culture setup in which airway epithelial cells are exposed to air on one side and medium on the other side.

  Aliases: air-interface, ALI culture

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• airway diseases

  disease

  Diseases of the airways for which airway epithelial metabolism is discussed as a potential research target.

  Aliases: airway disease

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• apical and basolateral feeding

  culture condition

  A culture condition in which medium is supplied to both the apical and basolateral sides of the epithelial layer.

  Aliases: apical plus basolateral feeding

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• engineered insert

  method, device

  A custom insert adapted for use in the Seahorse XF24 Analyzer to support metabolic measurements of airway epithelial cells at an air-liquid interface.

  Aliases: insert

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• extracellular acidification rate (ecar)

  measurement

  A readout of glycolytic activity measured in this assay.

  Aliases: ECAR, extracellular acidification rate

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• oxidative metabolism

  metabolic process

  Cellular energy metabolism dependent on oxygen-consuming mitochondrial respiration.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• oxygen consumption rate (ocr)

  measurement

  A readout of cellular respiration measured in this assay.

  Aliases: OCR, oxygen consumption rate

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• seahorse xf24 analyzer

  instrument

  A metabolic analyzer used here to measure cellular respiration and glycolysis in cultured airway epithelial cells.

  Aliases: XF24 Analyzer, Seahorse XF24

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review

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