Abstract
The deterioration of whole blood ex vivo represents a logistical hurdle in clinical and research settings. Here, a cocktail preservative is described that stabilizes leukocyte viability and erythrocyte morphology in whole blood under ambient storage. Neutrophil biostabilization was explored using a sophisticated microfluidic assay to examine the effectiveness of caspase inhibition to stabilize purified neutrophils. Following 72 h ambient storage, neutrophils remained fully functional to migrate towards chemical cues and maintained their ability to undergo NETosis after stimulation. Furthermore, stored neutrophils exhibited improved CD45 biomarker retention and reduced apoptosis and mortality compared to untreated controls. To stabilize erythrocyte morphology, a preservative solution was formulated using Taguchi methods of experimental design, and combined with the caspase inhibitor to form a whole blood cocktail solution, CSWB. CSWB was evaluated in blood from healthy donors and from women with metastatic breast cancer stored under ambient conditions for 72 h. CSWB-treated samples showed a significant improvement in erythrocyte morphology compared to untreated controls. Leukocytes in CSWB-treated blood exhibited significantly higher viability and CD45 biomarker retention compared to untreated controls. This 72 h shelf life under ambient conditions represents an opportunity to transport isolates or simply ease experimental timelines where blood degradation is problematic.
Highlights
The emergence of cutting edge clinical and research technologies has created a significant demand for the biostabilization of blood during transportation and storage[1]
To develop a preservative for erythrocytes and leukocytes in whole blood, we first characterized their deterioration under ambient storage conditions
Erythrocyte quality was assessed based on echinocyte enumeration
Summary
The emergence of cutting edge clinical and research technologies has created a significant demand for the biostabilization of blood during transportation and storage[1]. The ideal blood preservative should extend the shelf life of all cells in the blood for at least 72 h, sufficient time to transport samples globally and eliminate the need for immediate sample analysis This timeframe would enable sample storage over the weekend, easing experimental timelines. In modern blood banking, whole blood is leukoreduced within 24 h, followed by refrigeration in a cold storage solution to delay cellular damage[15] Several such preservatives have been developed including the FDA-licensed AS-1, AS-3 and AS-5 commercially available solutions[16, 17]. Component concentrations vary but typically consist of nutrients including NaCl, phosphate buffer system, adenine, dextrose and mannitol As these additives have been selected for erythrocytes, their formulations are not adequate for the storage of all blood components or ambient temperature, but serve as an excellent starting point for optimization. CSWB has significant applications for medical and diagnostic technologies using peripheral blood
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