Cryopreserved Peripheral Blood Mononuclear Cells Recover Mitochondrial Function Following Overnight Culture Revealed by High-Resolution Respirometry

Overnight recovery restores mitochondrial oxidative phosphorylation and maximal uncoupled respiration in cryopreserved peripheral blood mononuclear cells (PBMCs) to closely match freshly isolated cells.

• Why it matters: Fresh PBMCs enable mitochondrial respirometry, but limited flexibility and batching constrain large-scale or retrospective human studies.
• What they did: PBMCs from healthy young adults were tested as freshly isolated, directly thawed after cryopreservation, or cryopreserved with overnight culture recovery, then analyzed by Oroboros O2k respirometry after permeabilisation and substrate uncoupler inhibitor titration.
• The result: Direct thawing reduced ATP-coupled and maximal respiratory flux versus fresh cells, while overnight recovery substantially restored mitochondrial function with comparable viability, supporting cryopreserved PBMC use with a recovery step.

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