Abstract：
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.
摘要：
高分辨率呼吸测定（HRR）可以评估外周血单核细胞（PBMC）的生物能量，但不存在用于PBMC制备和HRR分析的标准化培养基。这里,我们研究了四种不同培养基(MiR05，PBS，RPMI,Plasmax)在计数上，尺寸,和完整PBMC的HRR分析(氧化图-O2k)。值得注意的是,当PBMC重悬于MiR05时，细胞计数比PBS或Plasmax高21%，由于固有的调整，导致HRR期间O2通量低估。此外，在MiR05中观察到较小的细胞大小和细胞聚集。根据我们的发现,我们建议Plasmax，PBS或RPMI比MiR05更适合用于完整PBMC的HRR分析。我们提供了Plasmax和PBS的氧溶解度因子，并鼓励进一步优化完整PBMC的标准化HRR方案。
