RT Journal Article
SR Electronic
T1 Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.01.10.475633
DO 10.1101/2022.01.10.475633
A1 Ida Bager Christensen
A1 Lucas Ribas
A1 Steen Larsen
A1 Flemming Dela
A1 Linn Gillberg
YR 2022
UL http://biorxiv.org/content/early/2022/01/11/2022.01.10.475633.abstract
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.Competing Interest StatementThe authors have declared no competing interest.