Preparation of Leukocyte-Poor Platelet Concentrates from Buffy Coats

Abstract

To study the influence of contaminating leukocytes on the storage conditions of platelet concentrates (PC), various amounts of leukocytes were added to identical PC. From 12 blood donations, 12 leukocyte-poor PC were prepared and pooled. Subsequently, the pool was divided into 12 identical PC. The plasma volume of the PC was 58.6±0.6 ml, the platelet concentration was 1.01±0.04x10^9/ml (mean ± SD) and the red cell contamination did not exceed 10^7 per PC. To 4 groups of 3 PC, pooled leukocytes were added from the same 12 blood donations. The leukocyte contamination for each group of 3 PC was 0.14±0.05, 1.96±0.09, 5.53±0.98 and 13.0±0.93x10^6/ml (mean ± SD) for groups I-IV, respectively. The PC were stored for 7 days at 22 °C in normal polyvinylchloride bags. A significant correlation was found between increasing concentrations of leukocytes in the PC and the drop in pH (r=-0.93), glucose consumption (r=-0.91), lactic acid production (r=0.93) and release of lactate dehydrogenase (r=0.92) during storage of the PC. The excretion of β-thromboglobulin, depletion of platelet adenine nucleotides, decreased ability to incorporate 3H-adenosine into metabolic nucleotides and poor morphology of the platelets were also significantly correlated with an increased number of leukocytes in the PC. These data show that high concentrations of leukocytes in PC have a significant detrimental effect on the viability of platelets during storage at 22 °C. We conclude that for good storage conditions of PC, the upper limit of leukocytes per PC should not exceed 10^7.

Factors influencing post-transfusional platelet increment in pediatric patients given hematopoietic stem cell transplantation

“…The platelet-rich plasma-derived PCs (PRP-PCs) were prepared by the method of Slichter and Harker, 17 while the buffy coatderived PCs (BC-PCs) were prepared using the method of Pietersz et al 18 The mean platelet content was respectively 70 ± 10.1 × 10 9 /U, and 66 ± 15.1 × 10 9 /U. The white cell contamination was 46 ± 30 × 10 6 /U in the BC-PCs, and about one log higher in the PRP-PCs.…”

“…Anti-HLA and platelet-specific antibodies were investigated before the conditioning regimen, at recovery from bone marrow aplasia following HSCT (days [15][16][17][18][19][20][21][22][23][24][25] and 40 days after HSCT. At the time of each platelet transfusion we also evaluated: body temperature (dichotomized as р37°C or Ͼ37°C), cytomegalovirus infection (defined as expression of the pp65 viral phosphoprotein on the leukocyte surface), 19 presence and severity of acute GVHD (classified according to Glucksberg et al 20 ), occurrence of thrombotic thrombocytopenic purpura (defined as microangiopathic hemolytic anemia with fragmentation of more than 5% of erythrocytes and no evidence of disseminated intravascular coagulation), occurrence of active mucosal bleeding (ie hematuria, blood in the stool, gum bleeding or epistaxis), spleen size (less or more than 1 cm below costal margin) or splenectomy before transplantation.…”

Preparation of Leukocyte-Poor Platelet Concentrates from Buffy Coats

“…For the Biotest and Terumo buffy coat bags (300 ml) the second centrifugation was at 22 "C for 6 min at 1,250 rpm (380 g; 2,150^ min) in the special inserts for the buckets of the unshielded 6-place J S 2.9 rotor of the Beckmann J6M, as previously described [11]. For the NPBI buffy coat bags (100 ml) different inserts were used for the cups of the shielded 6-place 4.2 rotor [1].…”

“…The optimal centrifugation conditions were 5 min at 1,200 rpm (250g; l,240g min). For all brands of bag the platelet-rich plasma was gently transferred into the bag that originally contained the SAG M solution [11], The bag with the residual buffy coat was then discarded.…”

Preparation of Leucocyte-Poor Platelet Concentrates from Buffy Coats

“…Leukocyte-poor platelet concentrates (lp-PC) can be prepared from buffy coats on a large scale by the appli cation of special inserts for centrifuge cups [1]. In 75% of routinely produced units, the lp-PC contain more than 55x 109 platelets and less than 107 leukocytes [1], In this method of lp-PC preparation many advantages of blood component manufacturing are combined.…”