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A novel model for studies of blood-mediated long-term responses to cellular transplants
Department of Immunology, Genetics and Pathology, Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden; Center for Clinical Research Dalarna-Uppsala University, Falun, Sweden .ORCID iD: 0000-0002-3671-5046
Department of Immunology, Genetics and Pathology, Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden .
Department of Immunology, Genetics and Pathology, Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden .
Department of Immunology, Genetics and Pathology, Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden .
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2015 (English)In: Upsala Journal of Medical Sciences, ISSN 0300-9734, E-ISSN 2000-1967, Vol. 120, no 1, p. 28-39Article in journal (Refereed) Published
Abstract [en]

Aims

Interaction between blood and bio-surfaces is important in many medical fields. With the aim of studying blood-mediated reactions to cellular transplants, we developed a whole-blood model for incubation of small volumes for up to 48 h.

Methods

Heparinized polyvinyl chloride tubing was cut in suitable lengths and sealed to create small bags. Multiple bags, with fresh venous blood, were incubated attached to a rotating wheel at 37°C. Physiological variables in blood were monitored: glucose, blood gases, mono- and divalent cations and chloride ions, osmolality, coagulation (platelet consumption, thrombin-antithrombin complexes (TAT)), and complement activation (C3a and SC5b-9), haemolysis, and leukocyte viability.

Results

Basic glucose consumption was high. Glucose depletion resulted in successive elevation of extracellular potassium, while sodium and calcium ions decreased due to inhibition of energy-requiring ion pumps. Addition of glucose improved ion balance but led to metabolic acidosis. To maintain a balanced physiological environment beyond 6 h, glucose and sodium hydrogen carbonate were added regularly based on analyses of glucose, pH, ions, and osmotic pressure. With these additives haemolysis was prevented for up to 72 h and leukocyte viability better preserved. Despite using non-heparinized blood, coagulation and complement activation were lower during long-term incubations compared with addition of thromboplastin and collagen.

Conclusion

A novel whole-blood model for studies of blood-mediated responses to a cellular transplant is presented allowing extended observations for up to 48 h and highlights the importance of stringent evaluations and adjustment of physiological conditions.

Place, publisher, year, edition, pages
Upsala Medical Society, 2015. Vol. 120, no 1, p. 28-39
National Category
Immunology in the medical area Clinical Laboratory Medicine
Identifiers
URN: urn:nbn:se:oru:diva-111681DOI: 10.3109/03009734.2014.965290ISI: 000350984700004PubMedID: 25322825Scopus ID: 2-s2.0-84924407569OAI: oai:DiVA.org:oru-111681DiVA, id: diva2:1838793
Funder
Swedish Research CouncilErnfors FoundationSwedish Diabetes AssociationInsamlingsstiftelsen Diabetes WellnessAFA InsuranceEU, FP7, Seventh Framework ProgrammeRegion DalarnaNIH (National Institutes of Health)
Note

This study was supported by grantsfrom the Swedish Medical Research Council, theNordic Insulin Fund, the Ernfors Family Fund, theSwedish Diabetes Association, Diabetes Wellness–Sweden, the Juvenile Diabetes Research FoundationInternational, AFA Insurances, and European Com-munity’s Seventh Framework Program. M.H. wassupported by grants from the Centre for ClinicalResearch (CKF) Dalarna, and Dalarna CountyCouncil. O.K.’s and B.N.’s positions are in partsupported by the National Institutes of Health.

Available from: 2024-02-19 Created: 2024-02-19 Last updated: 2024-02-20Bibliographically approved

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Hårdstedt, MariaLindblom, SusanneHong, Jaan

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