A novel model for studies of blood-mediated long-term responses to cellular transplants.

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.

• Authors: Maria Hårdstedt, Susanne Lindblom, Jaan Hong, Bo Nilsson, Olle Korsgren, Gunnar Ronquist
• Journal: Upsala journal of medical sciences (Ups J Med Sci) Vol. 120 Issue 1 Pg. 28-39 (Mar 2015) ISSN: 2000-1967 [Electronic] Sweden
• PMID: 25322825 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Blood Glucose
• Cations
• Chlorides
• Ions
• antithrombin III-protease complex
• Carbon Dioxide
• Antithrombin III
• Collagen
• Thromboplastin
• Peptide Hydrolases
• Glucose
• Oxygen

Topics

• Animals
• Antithrombin III (chemistry)
• Blood Gas Analysis
• Blood Glucose (metabolism)
• Carbon Dioxide (chemistry)
• Cations
• Cell Survival
• Cell Transplantation (methods)
• Chlorides (chemistry)
• Collagen (chemistry)
• Complement Activation
• Glucose (chemistry)
• Hemolysis
• Humans
• Hydrogen-Ion Concentration
• Ions
• Islets of Langerhans (cytology)
• Islets of Langerhans Transplantation (methods)
• Leukocytes (cytology)
• Osmolar Concentration
• Oxygen (chemistry)
• Pancreas (metabolism)
• Peptide Hydrolases (chemistry)
• Rabbits
• Thromboplastin (chemistry)
• Time Factors