Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis
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Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis. / Bager Christensen, Ida; Ribas, Lucas; Mosshammer, Maria; Abrahamsen, Marie Louise; Kühl, Michael; Larsen, Steen; Dela, Flemming; Gillberg, Linn.
I: Mitochondrion, Bind 77, 101890, 2024.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Choice of medium affects PBMC quantification, cell size, and downstream respiratory analysis
AU - Bager Christensen, Ida
AU - Ribas, Lucas
AU - Mosshammer, Maria
AU - Abrahamsen, Marie Louise
AU - Kühl, Michael
AU - Larsen, Steen
AU - Dela, Flemming
AU - Gillberg, Linn
N1 - Publisher Copyright: © 2024 The Author(s)
PY - 2024
Y1 - 2024
N2 - 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.
AB - 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.
KW - Cell count
KW - High-resolution respirometry
KW - Medium
KW - Oxygen solubility
KW - Peripheral blood mononuclear cells
U2 - 10.1016/j.mito.2024.101890
DO - 10.1016/j.mito.2024.101890
M3 - Journal article
C2 - 38718898
AN - SCOPUS:85193636163
VL - 77
JO - Mitochondrion
JF - Mitochondrion
SN - 1567-7249
M1 - 101890
ER -
ID: 393507299