Effective storage of granulocytes collected by centrifugation leukapheresis from donors stimulated with granulocyte-colony-stimulating factor

BACKGROUND: Donor stimulation with granulocyte–colony‐stimulating factor (G‐CSF) has increased the number of neutrophils (PMNs) that can be collected for granulocyte transfusion therapy. Clinical utility, however, has been limited by the inability to store functional PMNs ex vivo. This study was con...

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Published in:Transfusion (Philadelphia, Pa.) Pa.), 2005-12, Vol.45 (12), p.1876-1889
Main Authors: Hubel, Kai, Rodger, Elin, Gaviria, J. Milton, Price, Thomas H., Dale, David C., Liles, W. Conrad
Format: Article
Language:English
Subjects:
Adult
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Bacteria - immunology
Biological and medical sciences
Blood and lymphatic vessels
Blood Donors
Blood Preservation - methods
Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis
Bone marrow, stem cells transplantation. Graft versus host reaction
Cardiology. Vascular system
Cell Movement - immunology
Centrifugation
Cryopreservation - methods
Cytokines - metabolism
Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous
Female
Fungi - immunology
Granulocyte Colony-Stimulating Factor - administration & dosage
Granulocytes - cytology
Granulocytes - immunology
Granulocytes - metabolism
Humans
Indicators and Reagents
Injections, Subcutaneous
Leukapheresis - methods
Leukocyte Count
Leukocyte Transfusion - adverse effects
Luminol
Male
Medical sciences
Middle Aged
Neutrophils - cytology
Neutrophils - immunology
Respiratory Burst - immunology
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
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Summary:BACKGROUND: Donor stimulation with granulocyte–colony‐stimulating factor (G‐CSF) has increased the number of neutrophils (PMNs) that can be collected for granulocyte transfusion therapy. Clinical utility, however, has been limited by the inability to store functional PMNs ex vivo. This study was conducted to determine whether granulocyte products from G‐CSF–stimulated donors could be effectively stored at reduced temperature (22°C vs. 10°C) with maintenance of functional properties in vitro and in vivo. STUDY DESIGN AND METHODS: Nine normal subjects received G‐CSF (600 µg subcutaneously) 12 hours before centrifugation leukapheresis. Granulocyte products were divided and stored for 24 and 48 hours under four conditions: 1) 22°C; 2) 22°C, with supplemental G‐CSF (100 ng/mL); 3) 10°C; and 4) 10°C, with supplemental G‐CSF. Functional PMN activity during ex vivo storage was assessed in vitro and in vivo by the skin‐window technique for granulocytes stored at 10°C for 24 hours. RESULTS: Surface expression of CD11b/CD18, CD14, CD16, CD32, and CD64 was maintained during 48‐hour storage at reduced temperature. Inducible respiratory burst activity, bactericidal activity, and fungicidal activity were preserved during storage for 48‐hour storage at 10°C. Proinflammatory cytokine production was decreased in product stored at 10°C. Supplemental G‐CSF ex vivo did not substantially improve functional activity during storage. After storage at 10°C for 24 hours, in vitro chemotactic potential was maintained, and transfused granulocytes retained capacity to circulate and migrate appropriately in vivo. CONCLUSIONS: Granulocyte product collected by centrifugation leukapheresis from G‐CSF–stimulated donors can be effectively stored at subphysiologic temperature for 24 hours with preservation of functional activity. Storage at 10°C appears to be slightly superior to storage at 22°C.
ISSN:0041-1132
1537-2995
DOI:10.1111/j.1537-2995.2005.00636.x