Page 205 - 2020_08-Haematologica-web

Metabolism of stored human and macaque RBC
fresh and stored RM RBC had significantly higher levels of most phosphatidylserines (Online Supplementary Figure S6C-E) and short/medium-chain, but not long and very- long chain, fatty acyl-phosphatidylethanolamines (Online Supplementary Figure S6F, G).
Validation using targeted quantitative metabolomics and lipidomics
The initial, global approach generated extensive, but relative, quantification data for several pathways. To con- firm critical observations, we performed validation exper-
A
iments on all RBC and supernatant samples using target- ed quantitative methods with stable isotope-labeled internal standards (Figure 7A). Results are reported in tab- ular and vectorial formats (Online Supplementary Table S2 and Online Supplementary Figure S7). There was a substan- tial overlap (r2 >0.75) between relative and absolute val- ues for critical metabolites in amino acid, redox (glu- tathione and purine oxidation), and fatty acid metabolic pathways, which demonstrated significant differences between the two species as a function of storage duration (Figure 7B-F) and sex (Online Supplementary Figure S4E-G).
Figure 5. Species- and storage time-specific metabolic changes in Rhesus macaque and human red blood cells and supernatants: focus on purine metabolism, urea cycle, and carboxylate metabolism. (A, B) Data for Rhesus macaques are shown in green, those for humans are represented in red. Supernatant metabolites are shown in the right-hand side of panel (B), outside the representative lipid bilayer of the cellular membrane.
B
haematologica | 2020; 105(8)
2181