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Peer-reviewedCC BY-NCSTAR protocols

Protocol for isolation and cryopreservation of human peripheral blood mononuclear cells for high-resolution respirometry.

  • Alejandra Romero1
  • Alba Timón-Gómez2
  • Jaime R Willis3
  • Anja Perfler4
  • Theresa Lindlbauer5
  • Michael Knoflach5
  • Erich Gnaiger2
5 affiliations
  1. 1VASCage-Centre on Clinical Stroke Research, Innsbruck 6020, Austria. Electronic address: [email protected].
  2. 2Oroboros Instruments, Innsbruck 6020, Austria.
  3. 3Oroboros Instruments, Innsbruck 6020, Austria. Electronic address: [email protected].
  4. 4Department of Neurology, Medical University of Innsbruck, Innsbruck 6020, Austria.
  5. 5VASCage-Centre on Clinical Stroke Research, Innsbruck 6020, Austria.
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Abstract

High-resolution respirometry (HRR) of peripheral blood mononuclear cells (PBMCs) relies on controlled preanalytical procedures. Here, we present an optimized workflow for PBMC isolation from human whole blood, followed by short-term cryopreservation at -80°C. The protocol includes rapid thawing to preserve cell viability, washing steps to minimize platelet contamination, and quality-control assessment of PBMC number, viability, and composition before HRR measurements. Critical handling steps are optimized and validated by HRR against freshly prepared cells.

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Cite

BibTeXRIS
BibTeX
@article{romero2026protocol,
  title = {{Protocol for isolation and cryopreservation of human peripheral blood mononuclear cells for high-resolution respirometry.}},
  author = {Alejandra Romero and Alba Timón-Gómez and Jaime R Willis and Anja Perfler and Theresa Lindlbauer and Michael Knoflach and Erich Gnaiger},
  journal = {STAR protocols},
  year = {2026},
  month = oct,
  doi = {10.1016/j.xpro.2026.104881},
  url = {https://europepmc.org/article/MED/42842376},
  note = {CC BY-NC},
}
RIS
TY  - JOUR
TI  - Protocol for isolation and cryopreservation of human peripheral blood mononuclear cells for high-resolution respirometry.
AU  - Alejandra Romero
AU  - Alba Timón-Gómez
AU  - Jaime R Willis
AU  - Anja Perfler
AU  - Theresa Lindlbauer
AU  - Michael Knoflach
AU  - Erich Gnaiger
PY  - 2026
DA  - 2026/10/07
JO  - STAR protocols
DO  - 10.1016/j.xpro.2026.104881
UR  - https://europepmc.org/article/MED/42842376
AB  - High-resolution respirometry (HRR) of peripheral blood mononuclear cells (PBMCs) relies on controlled preanalytical procedures. Here, we present an optimized workflow for PBMC isolation from human whole blood, followed by short-term cryopreservation at -80°C. The protocol includes rapid thawing to preserve cell viability, washing steps to minimize platelet contamination, and quality-control assessment of PBMC number, viability, and composition before HRR measurements. Critical handling steps are optimized and validated by HRR against freshly prepared cells.
SN  - 2666-1667
ER  -