Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis
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.
2019 ◽
Vol 220
(1)
◽
pp. S93
Keyword(s):
2020 ◽
Vol 11
(2)
◽
pp. 61-67
Keyword(s):
1967 ◽
Vol 31
◽
pp. 45-46
Keyword(s):
1994 ◽
Vol 144
◽
pp. 593-596
1994 ◽
Vol 144
◽
pp. 541-547
1970 ◽
Vol 28
◽
pp. 260-261
Keyword(s):
1970 ◽
Vol 28
◽
pp. 258-259