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Related Experiment Video

Updated: May 12, 2026

Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes
08:04

Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes

Published on: August 16, 2021

Assessing Mitochondrial Respiratory Complex-Associated Function From Previously Frozen Mouse Placental Tissue.

Tina Podinic1, Donald Xhuti1, Cristina Monaco1

  • 1Department of Pediatrics and the Graduate Program in Medical Sciences, McMaster University, Hamilton, ON, Canada.

Bio-Protocol
|May 11, 2026
PubMed
Summary

Researchers developed a new Seahorse XF method to measure mitochondrial respiration in frozen placental tissue. This approach enables functional analysis of placental mitochondria, crucial for understanding fetal development and environmental exposures.

Keywords:
BioenergeticsElectron transport-chain enzyme activityFresh tissueFrozen tissueMetabolismMitochondriaOxygen consumptionPlacentaRespirationRespirometry

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Last Updated: May 12, 2026

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Area of Science:

  • Mitochondrial biology
  • Placental research
  • Biochemistry

Background:

  • Placental mitochondrial activity is vital for fetal growth and nutrient transport.
  • Dysfunctional mitochondria are linked to placental dysfunction.
  • Current respirometry methods are impractical for large-scale studies due to tissue requirements.

Purpose of the Study:

  • To validate a Seahorse XF protocol for measuring mitochondrial respiration in frozen placental tissue.
  • To enable functional interrogation of placental mitochondria in large-scale prenatal studies.
  • To provide a streamlined method for assessing respiratory enzyme complex function.

Main Methods:

  • Developed and validated a Seahorse XF approach for frozen placental tissue.
  • Restored depleted matrix substrates lost during freeze-thaw cycles.
  • Assessed complex I and II-associated respiration using the Seahorse XFe24 Analyzer.

Main Results:

  • Demonstrated comparable oxygen consumption readouts between fresh and frozen placentae.
  • Identified distinct differences in oxygen consumption between fresh and frozen placentae without exogenous NADH.
  • Established a simplified protocol for assessing respiratory enzyme complex-associated respiration.

Conclusions:

  • The described Seahorse XF protocol effectively measures mitochondrial respiration in previously frozen placental tissue.
  • This method facilitates functional analysis of archived placental samples, supporting large-scale prenatal studies.
  • The protocol is convenient, streamlined, and yields data within 4 hours.