Rethinking mitochondrial isolation for the next generation of mitochondrial therapies.
- 1Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada.
- 2Department of Biomedical Engineering, University of Alberta, Edmonton, AB, Canada.
- 3Canadian Blood Services Innovation, Canadian Blood Services, Edmonton, AB, Canada.
Abstract
Mitochondrial isolation is fundamental to the study of mitochondrial biology and is increasingly relevant to emerging therapeutic applications such as mitochondrial transplantation. However, isolation procedures can substantially influence mitochondrial yield, purity, structural integrity, and bioenergetic function, making method selection an important determinant of experimental and translational outcomes. This review examines current mitochondrial isolation strategies, including physical disruption, chemical permeabilization, emerging microfluidic and automated dissociation technologies, and downstream separation approaches such as differential centrifugation, density-gradient centrifugation, filtration, and immunoaffinity enrichment. Each approach offers distinct advantages and limitations depending on the biological source, processing requirements, and intended application. Conventional methods remain widely used because of their accessibility and flexibility, but may introduce operator variability, mechanical stress, contamination, or loss of mitochondrial function. More selective purification strategies can improve sample purity, although often at the cost of additional processing time, technical complexity, and reduced recovery. Emerging technologies, including standardized mechanical dissociation and microfluidic disruption, may provide greater control over mitochondrial release, but require broader validation across tissues and sample volumes. A persistent challenge across isolation workflows is balancing mitochondrial recovery with preservation of organelle integrity and function. Future progress will require standardized, scalable isolation strategies that consistently preserve mitochondrial quality across different biological sources and downstream applications. Continued refinement of these approaches will be essential for advancing mitochondrial research and supporting the development of mitochondrial-based therapies.
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BibTeX
@article{simpson2026rethinking,
title = {{Rethinking mitochondrial isolation for the next generation of mitochondrial therapies.}},
author = {Ethan A Simpson and Celina Phan and Jason P Acker},
journal = {Frontiers in physiology},
year = {2026},
month = oct,
doi = {10.3389/fphys.2026.1975105},
url = {https://europepmc.org/article/MED/42903348},
note = {CC BY},
}
RIS
TY - JOUR
TI - Rethinking mitochondrial isolation for the next generation of mitochondrial therapies.
AU - Ethan A Simpson
AU - Celina Phan
AU - Jason P Acker
PY - 2026
DA - 2026/10/06
JO - Frontiers in physiology
DO - 10.3389/fphys.2026.1975105
UR - https://europepmc.org/article/MED/42903348
AB - Mitochondrial isolation is fundamental to the study of mitochondrial biology and is increasingly relevant to emerging therapeutic applications such as mitochondrial transplantation. However, isolation procedures can substantially influence mitochondrial yield, purity, structural integrity, and bioenergetic function, making method selection an important determinant of experimental and translational outcomes. This review examines current mitochondrial isolation strategies, including physical disruption, chemical permeabilization, emerging microfluidic and automated dissociation technologies, and downstream separation approaches such as differential centrifugation, density-gradient centrifugation, filtration, and immunoaffinity enrichment. Each approach offers distinct advantages and limitations depending on the biological source, processing requirements, and intended application. Conventional methods remain widely used because of their accessibility and flexibility, but may introduce operator variability, mechanical stress, contamination, or loss of mitochondrial function. More selective purification strategies can improve sample purity, although often at the cost of additional processing time, technical complexity, and reduced recovery. Emerging technologies, including standardized mechanical dissociation and microfluidic disruption, may provide greater control over mitochondrial release, but require broader validation across tissues and sample volumes. A persistent challenge across isolation workflows is balancing mitochondrial recovery with preservation of organelle integrity and function. Future progress will require standardized, scalable isolation strategies that consistently preserve mitochondrial quality across different biological sources and downstream applications. Continued refinement of these approaches will be essential for advancing mitochondrial research and supporting the development of mitochondrial-based therapies.
SN - 1664-042X
ER -