Abstract
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.
Highlights
Several of the achievements in modern medicine, e.g. haemodialysis, joint and dental prostheses, vessel grafts, vascular stents, and organ and cellular transplants, involve contact between circulating human blood and biomaterials, cells, or tissues
Defined in islet transplantation, instant blood-mediated inflammatory reaction (IBMIR) has later been reported for hepatocytes, mesenchymal stromal cells (MSCs), and recently in autologous islet transplantation [13,14,15]
We describe the changes in blood physiology during incubations for up to 72 h and evaluate the relative importance of these changes and ways to counteract them
Summary
Several of the achievements in modern medicine, e.g. haemodialysis, joint and dental prostheses, vessel grafts, vascular stents, and organ and cellular transplants, involve contact between circulating human blood and biomaterials, cells, or tissues. With regard to transplantation of functional pancreatic islets, in vitro blood models have been crucial in describing the instant blood-mediated inflammatory reaction (IBMIR) following infusion of ABO-compatible donor islets into the portal vein of the recipient [3,10]. This reaction starts with an immediate thrombotic and inflammatory reaction with activation of complement and coagulation cascades together with platelet aggregation, and is believed to cause a deleterious rapid loss of transplanted islets prior to engraftment [3,11,12]. This demands large volumes of valuable cellular material and drugs to be tested and makes it impossible to run multiple treatment groups and replicates during the same experiment
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