Ex Vivo Assessment of Extracellular Acidification Rate in Murine Intestinal Tissue

Area of Science:

• Cellular and Molecular Metabolism
• Physiology
• Biochemistry

Background:

• Seahorse assays are standard for measuring cellular glycolysis and mitochondrial function in vitro.
• Assessing real-time metabolism in ex vivo tissue samples using Seahorse assays is not yet standardized.
• Existing methods primarily focus on adherent or suspension cell cultures.

Purpose of the Study:

• To develop and present a protocol for assessing glycolysis in ex vivo murine intestinal samples using Seahorse metabolic assays.
• To adapt Seahorse assays for functional metabolic measurements in tissue samples.
• To complement existing in vitro metabolic assessment methods with ex vivo tissue analysis.

Main Methods:

• Utilized the Agilent Seahorse XFe24 platform with Islet Capture microplates.
• Measured extracellular acidification rate (ECAR) to assess glycolysis.
• Applied the protocol to ex vivo murine intestinal tissue samples.

Main Results:

• Successfully established a protocol for measuring glycolytic function in ex vivo intestinal tissue.
• Demonstrated the applicability of Seahorse assays to tissue samples for functional metabolism studies.
• Provided a method to obtain real-time metabolic data from tissues, complementing other molecular data.

Conclusions:

• This protocol enables functional assessment of organ-level metabolism in ex vivo tissue samples.
• The method can be adapted to measure mitochondrial respiration (oxygen consumption rate, OCR) in various tissue types.
• Expands the utility of Seahorse assays for interrogating systemic nutrient metabolism and physiology.