Abstract
The rapid degradation of blood ex vivo imposes logistical limitations on the utilization of blood-borne cells in medical diagnostics and scientific investigations. A fundamental but overlooked aspect in the storage of this fluid tissue is blood settling, which induces physical stress and compaction, aggregates blood cells, and causes collateral damage due to leukocyte activation. Here we show that the polymer Ficoll 70 kDa stabilized blood samples and prevented blood settling over the course of 72 hours, primarily by inhibiting depletion-mediated red blood cell aggregation. Physical stabilization decreased echinocyte formation, improved leukocyte viability, and inhibited the release of neutrophil elastase—a marker of neutrophil extracellular trap formation. In addition, Ficoll-stabilized blood was compatible with common leukocyte enrichment techniques including red blood cell lysis and immunomagnetic purification. This study showed for the first time that blood settling can be prevented using polymers and has implications in diagnostics.
Highlights
Peripheral blood is the most frequently accessed tissue in the clinic, and the isolation of blood-borne cells is of broad clinical and scientific importance in hematology, transfusion, immunology, regenerative medicine, and oncology[1,2,3,4]
Note that these images do not represent the entire length of the erythrocyte settling rate (ESR) assay. (B) ESR of blood samples in whole blood (WB) and blood containing different concentrations of Ficoll 70 kDa (F70) over 72 hours (WB, n = 11; 0% F70, n = 9; 5% F70, n = 8; 10% F70, n = 11; 15% F70, n = 9). 0% F70 represents the control condition which was treated with RPMI medium without F70
The addition of 5%, 10%, and 15% F70 greatly decreased the ESR at all measured timepoints (Fig. 1B; p < 0.0001 compared to WB at 24, 48, and 72 hours)
Summary
Peripheral blood is the most frequently accessed tissue in the clinic, and the isolation of blood-borne cells is of broad clinical and scientific importance in hematology, transfusion, immunology, regenerative medicine, and oncology[1,2,3,4]. Given that many clinically relevant assays such as sequencing and expression profiling are best performed in large medical centers or diagnostic laboratories, the logistical needs of blood storage and transportation impose severe limitations on the dissemination of next-generation blood-based medical technologies. A fundamental aspect in the storage of this fluid tissue, blood cell settling, has been largely overlooked. This physical event alters cellular activities and leads to platelet aggregation[14,15] and functional deterioration of leukocytes[16,17]. Note that these images do not represent the entire length of the ESR assay. The ability to suspend blood in a homogeneous phase may improve the preservation of WB ex vivo and facilitate the logistics of off-site or delayed processing of samples
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