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Purified Granulocyte Concentrates from Buffy Coats with Extended Storage Time.

Gerd Klinkmann1,2, Fanny Doss3, Sandra Doss2,3

  • 1Department of Anesthesiology, Intensive Care Medicine and Pain Therapy, University Medical Center, Rostock, Germany.

Transfusion Medicine and Hemotherapy : Offizielles Organ Der Deutschen Gesellschaft Fur Transfusionsmedizin Und Immunhamatologie
|December 12, 2024
PubMed
Summary

Pooled granulocyte concentrates (GCs) from buffy coats offer an alternative to apheresis GCs. Reducing red blood cell and platelet contamination extends GC shelf life to 72 hours, improving availability for patients.

Keywords:
Buffy coatGranulocyte concentrateTransfusionTransfusion medicine

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Area of Science:

  • Hematology
  • Transfusion Medicine
  • Cellular Therapy

Background:

  • Granulocyte concentrates (GCs) are typically prepared via single-donor apheresis.
  • Short shelf life (24h) of GCs is due to rapid pH decrease and cell death from red blood cell contamination.
  • Pooled GCs from whole blood donations could enhance product availability.

Purpose of the Study:

  • To develop a manufacturing process for pooled granulocyte concentrates (GCs).
  • To investigate the viability and functionality of pooled GCs over an extended storage period.
  • To assess the potential of buffy coat pooling for improving GC availability.

Main Methods:

  • Six ABO-identical buffy coats were pooled.
  • Hydroxyethyl starch induced spontaneous red blood cell sedimentation.
  • Leukocyte-rich supernatant underwent saline washes to reduce platelets and was resuspended in donor plasma.
  • Leukocyte concentrates were stored at 20-24°C for up to 72 hours.
  • Evaluated cell counts, viability, pH, blood gases, phagocytosis, and oxidative burst activity.

Main Results:

  • Red blood cell and platelet contamination reduced by over 90% (to 0.4% and 6.1% respectively).
  • Approximately 50% of original leukocytes were extracted.
  • White blood cell counts remained stable over 72 hours of storage (p=0.12).
  • Viability exceeded 98% throughout the storage period.
  • Granulocyte phagocytosis and oxidative burst activity remained above 95%.

Conclusions:

  • Pooled buffy coat GCs represent a viable alternative to apheresis-derived GCs.
  • Significant reduction in RBC and platelet contamination extends GC shelf life to 72 hours.
  • Therapeutic doses require 15-20 buffy coats for at least 1x10^10 granulocytes.