Stiffness and ATP recovery of stored red blood cells in serum

Abstract

In transfusion medicine, there has been a decades-long debate about whether the age of stored red blood cells (RBCs) is a factor in transfusion efficacy. Existing clinical studies investigating whether older RBCs cause worse clinical outcomes have provided conflicting information: some have shown that older blood is less effective, while others have shown no such difference. The controversial results could have been biased by the vastly different conditions of the patients involved in the clinical studies; however, another source of inconsistency is a lack of understanding of how well and quickly stored RBCs can recover their key parameters, such as stiffness and ATP concentration, after transfusion. In this work, we quantitatively studied the stiffness and ATP recovery of stored RBCs in 37 °C human serum. The results showed that in 37 °C human serum, stored RBCs are able to recover their stiffness and ATP concentration to varying extents depending on how long they have been stored. Fresher RBCs (1–3 weeks old) were found to have a significantly higher capacity for stiffness and ATP recovery in human serum than older RBCs (4–6 weeks old). For instance, for 1-week-old RBCs, although the shear modulus before recovery was 1.6 times that of fresh RBCs, 97% of the cells recovered in human serum to have 1.1 times the shear modulus of fresh RBCs, and the ATP concentration of 1-week-old RBCs after recovery showed no difference from that of fresh RBCs. However, for 6-week-old RBCs, only ~70% of the RBCs showed stiffness recovery in human serum; their shear modulus after recovery was still 2.1 times that of fresh RBCs; and their ATP concentration after recovery was 25% lower than that of fresh RBCs. Our experiments also revealed that the processes of stiffness recovery and ATP recovery took place on the scale of tens of minutes. We hope that this study will trigger the next steps of comprehensively characterizing the recovery behaviors of stored RBCs (e.g., recovery of normal 2,3-DPG [2,3-Diphosphoglycerate]and SNO [S-nitrosation] levels) and quantifying the in vivo recovery of stored RBCs in transfusion medicine.

Cyclic mechanical stresses alter erythrocyte membrane composition and microstructure and trigger macrophage phagocytosis

“…Finally, to obtain the intracellular ATP concentration, the average measured concentration was normalized by the total number of cells lysed and the mean volume obtained in previous experiments. ATP measured concentrations were similar to previous studies52 .RBCs recirculation and crossingtime determination. Flow was driven through the device at a fix flow rate (100µl/h) using a peristaltic pump (IPC, ISMATEC) (Suppl Fig 1F); wider channels (5 m) were added at both sides of the microchannel array to ensure that microchannel clogging by stuck RBCs did not significantly affect the pressure difference across the array.…”

Deformability of Stored Red Blood Cells

“…And although there are conflicting data (Weed et al, 1969;Karger et al, 2012;Huisjes et al, 2018) on the relationship between intracellular ATP content and RBC deformability, some authors have tested the effectiveness of restoring the expected level of ATP in reducing PRBC stiffness. Thus, Xu et al (2019) examined whether stiffness of PRBCs can be restored in human serum (at temperatures below 37 • C) and the role of storage duration in this process. Authors demonstrated that PRBCs could recover their deformability and ATP concentration in human serum, with the extent of recovery decreasing as a function of storage duration (Xu et al, 2019).…”

“…Thus, Xu et al (2019) examined whether stiffness of PRBCs can be restored in human serum (at temperatures below 37 • C) and the role of storage duration in this process. Authors demonstrated that PRBCs could recover their deformability and ATP concentration in human serum, with the extent of recovery decreasing as a function of storage duration (Xu et al, 2019). Moreover, the recovery processes took place on a scale of tens (10 -90 min) of minutes, and in the case of a more extended storage period, a longer recovery was required.…”

Red Blood Cell Stiffness and Adhesion Are Species-Specific Properties Strongly Affected by Temperature and Medium Changes in Single Cell Force Spectroscopy

“…The RBC cytoskeleton deforms generally passively. Only one complex is actively triggered [ 54 ]. ATP production in RBCs depends on the availability of glucose as substrate and on the enzymatic activity of the Embden–Meyerhof pathway.…”