TY - UNPB
T1 - Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis
AU - Christensen, Ida Bager
AU - Ribas, Lucas
AU - Larsen, Steen
AU - Dela, Flemming
AU - Gillberg, Linn
PY - 2022
Y1 - 2022
N2 - High-resolution respirometry (HRR) can assess 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 quantification, size, and HRR analysis (Oxygraph-O2k) of intact PBMCs. Remarkably, PBMC quantification was 21% higher in MiR05 than PBS and Plasmax, and 28% higher than in RPMI, causing O2 flux underestimation during HRR due to inherent adjustments. Moreover, smaller cell size of PBMCs and aggregation was observed in MiR05. We suggest optimization of HRR with a standardized, plasma-like medium for future HRR analysis of intact PBMCs.
AB - High-resolution respirometry (HRR) can assess 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 quantification, size, and HRR analysis (Oxygraph-O2k) of intact PBMCs. Remarkably, PBMC quantification was 21% higher in MiR05 than PBS and Plasmax, and 28% higher than in RPMI, causing O2 flux underestimation during HRR due to inherent adjustments. Moreover, smaller cell size of PBMCs and aggregation was observed in MiR05. We suggest optimization of HRR with a standardized, plasma-like medium for future HRR analysis of intact PBMCs.
UR - https://doi.org/10.1101/2022.01.10.475633
U2 - 10.1101/2022.01.10.475633
DO - 10.1101/2022.01.10.475633
M3 - Preprint
VL - 2022
BT - Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis
PB - bioRxiv
ER -