Isolation and cryopreservation of human peripheral blood mononuclear cells for high-resolution respirometry

Version 2

Abstract

Ensuring high-quality preparations of peripheral blood mononuclear cells (PBMCs) is essential for obtaining consistent results matching the accuracy of high-resolution respirometry. Here, we present an advanced protocol to isolate and cryopreserve human PBMCs. We describe steps to maintain viability of cryopreserved cells and minimize platelet contamination through optimized centrifugation. This protocol provides a valuable reference for optimized PBMC isolation and cryopreservation in mitochondrial research and diagnostic applications.
This PBMC isolation and cryopreservation protocol is a modified and optimized version of a widely applied procedure ^1–4, and is tested by high-resolution respirometry with reference to freshly prepared cells.

Cite:

Romero A, Timón-Gómez A, Willis JR, Perfler A, Lindlbauer T, Knoflach M, Gnaiger E (2026) Isolation and cryopreservation of human peripheral blood mononuclear cells for high-resolution respirometry. BEC preprints 2026.2.v2. https://doi.org/10.26124/becprep.2026-0002.v2