Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis.

High-resolution respirometry (HRR) can assess peripheral blood mononuclear cell (PBMC) bioenergetics, but no standardized medium for PBMC preparation and HRR analysis exist. Here, we study the effect of four different media (MiR05, PBS, RPMI, Plasmax) on the count, size, and HRR (Oxygraph-O2k) of intact PBMCs. Remarkably, the cell count was 21 % higher when PBMCs were resuspended in MiR05 than in PBS or Plasmax, causing O2 flux underestimation during HRR due to inherent adjustments. Moreover, smaller cell size and cell aggregation was observed in MiR05. Based on our findings, we propose that Plasmax, PBS or RPMI is more suitable than MiR05 for HRR of intact PBMCs. We provide oxygen solubility factors for Plasmax and PBS and encourage further optimization of a standardized HRR protocol for intact PBMCs. Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.

Citation

Ida Bager Christensen, Lucas Ribas, Maria Mosshammer, Marie-Louise Abrahamsen, Michael Kühl, Steen Larsen, Flemming Dela, Linn Gillberg. Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis. Mitochondrion. 2024 Jul;77:101890

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PMID: 38718898

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