The preparation and validation of blood components has previously been described in detail [19 (link)]. Briefly, two units of whole blood (WB; 1 unit = 450 ml ± 10% v/v) were collected from each donor sheep into blood packs (Fresenius Hemocare NPBI), containing CPD.A1 anticoagulant. Components were separated by centrifugation and extraction using a Compomat G4 (Fresenius Hemocare) optimised for the separation of sheep blood components, to yield platelet-rich plasma, red cells and buffy coat. Platelet-rich plasma was further centrifuged to produce platelet-poor plasma and platelet concentrate. Leucodepletion of components was performed using in-line leucoreduction filters (Fresenius/NPBI).
Prior to transfusion, 10–20 ml of each component was sampled for analysis and archive storage/bioassay. For each component, the volume was estimated based on weight (adjusted for specific gravity). Numbers of leucocytes in non-leucodepleted and leucodepleted components were measured by flow cytometry, using a Leucocount kit (BD Biosciences). Numbers of platelets in platelet concentrates and plasma fractions were counted manually using a haemocytometer. In whole blood, red cells and buffy coat, the haematocrit was measured, using a haematocrit centrifuge and reader (Hawksley), to determine the distribution of plasma in different components.
Prior to transfusion, 10–20 ml of each component was sampled for analysis and archive storage/bioassay. For each component, the volume was estimated based on weight (adjusted for specific gravity). Numbers of leucocytes in non-leucodepleted and leucodepleted components were measured by flow cytometry, using a Leucocount kit (BD Biosciences). Numbers of platelets in platelet concentrates and plasma fractions were counted manually using a haemocytometer. In whole blood, red cells and buffy coat, the haematocrit was measured, using a haematocrit centrifuge and reader (Hawksley), to determine the distribution of plasma in different components.
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