Verlängerte Lagerungsmöglichket von Granulozytenkonzentraten aus gepoolten Buffy Coats durch Aufreinigung
Jens Altrichter (Rostock / DE), Sophie Brabant (Rostock / DE), Torsten J. Schulze (Springe / DE), Jessica Rach (Springe / DE), Fanny Doss (Rostock / DE), Sandra Doss (Rostock / DE), Gerd Klinkmann (Rostock / DE)
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JA, FD, SD sind Angestellte der ARTCLINE GmbH und Co-Erfinder von Patenten.
Die anderen Autoren haben keine Interessenkonflikte bezüglich dieser Studie.
Background:
In Germany, granulocyte concentrates are typically produced by apheresis and require donor pretreatment with G-CSF and dexamethasone, often resulting in time delays, limited availability and potential side effects for the donor.
In contrast, producing GCs from pooled buffy coats (BCs) offers expanded availability and fewer donor risks. Recently, we published the production of purified GCs using BCs and hydroxyethyl starch or apheresis GCs (1, 2). Here, we present a manufacturing process involving the pooling of BCs and purification based on gelatine sedimentation to achieve the desired cell counts and extended storage. Viability, functionality, and metabolism of the cells were investigated during prolonged storage.
Methods:
Up to 20 ABO blood group-identical BCs were pooled. After adding gelatine (Gelafundin), the red blood cells sedimented. The remaining leukocyte-rich supernatant was washed three times with saline to reduce platelet content and was resuspended in donor plasma. The purified pooled GC (ppGC) was stored at 20–24°C without agitation for up to 96 hours. Cell count and viability, metabolic parameters, and functionality were monitored daily.
Results:
Pooling of 12 BCs resulted in ppGC with 8.5E9 granulocytes (N=12, range 5.3-12.2E9) corresponding to the EDQM specifications for pooled granulocyte concentrates (at least 5E9). Pooling of 20 BCs resulted in ppGC with 1.22E10 granulocytes (N=6, range 1.03-1.40E10) corresponding to the German Hemotherapy specifications for granulocyte concentrates (at least 1E10). During storage for 96h viability and phagocytosis rate were above 80%.
Conclusion:
Granulocyte concentrates (GCs) from pooled buffy coats meet required granulocyte cell counts and demonstrate an extended storability. Both hydroxyethyl starch and gelatine can be used. This may allow offering granulocyte concentrates as an of the shelf product in the future for immediate use and broad applicability for transfusion and extracorporeal immune therapy (3).
Transfus Med Hemotherapy. 12. April 2024;1–10Transfusion (Paris). 2022;62:194–204Crit Care. 2011;15:R82