Investigation of how different filters affect some biochemical properties of stored platelet concentrates Jaremo P, Kutti J. Investigation of how different filters affect some biochemical properties of stored platelet concentrates. Eur J Haematol 1992: 49: 25-28. Abstract: The aim of the present study was to investigate how four different filters, i.e. Imugard IG500 (Terumo, Japan), Miropore (Miramed, Italy), Pall P1-100 (Pall, USA) and Sepacell Pl-1OA (Asahi, Japan) affect some biochemical properties of platelet concentrates. The work was conducted using 42 pairs of platelet Concentrates. After 2 days of storage, one of the preparations was filtered and the other served as an unfiltered control. Immediately after filtration, determination of the platelet count, desarginated activated complement factor 3 (C3a des arg) and the 'extracellular and total concentrations of platelet factor 4 (PF4) and lactate dehydrogenase (LDH) were carried out on both these platelet concentrates. After an additional storage period of 3 d, extracellular concentrations of PF4 and L D H were determined on both concentrates. A significant decrease of extracellular PF4 concentration was found immediately after filtration when Pall P1-100 and Imugard IG500 were used. During the 3-d storage after filtration, the concentrates filtered with Imugard IG500 and Pall P1-100 demonstrated significantly higher platelet lysis as compared to the unfiltered controls. It is concluded that the present work demonstrates storage instability after filtration with Imugard IG500 and Pall P1- 100. Therefore, platelet concentrates filtered with these filters would not appear to be suitable for storage.

Introduction

It has been established that the rise of antibodies against HLA antigens in patients receiving platelet transfusion therapy is delayed if the number of white blood cells in platelet concentrates is reduced by filtration prior to transfusion (1-3). With the development of new filters (4, 5 ) the filtration procedure has become simplified such that it can be performed while the patient receives platelet transfusion therapy. The present work investigates four different platelet filters with respect to their effects upon platelet concentrates both immediately after filtration and after an additional 3-d storage period following filtration. Desarginated activated complement factor 3 (C3a des arg) served as a marker for complement activation (6) while the extracellular fractions of lactate dehydrogenase (LDH) and platelet factor 4 (PF4) were used to estimate platelet lysis and activation (7, S), respectively.

P. Jaremo and J. Kutti * Department of Transfusion Medicine, Orebro Medical Center Hospital, Orebro, and * Department of Medicine Ostra Hospital, Gothenburg University, Gothenburg, Sweden

Key words: C3a des arg - platelets - platelet factor 4 - platelet filtration Correspondence: Petter Jaremo, MD, The blood Transfusion Center, Orebro Medical Center Hospital, S-701 85 Orebro Sweden Accepted for publication 21 April 1992

Methods Platelet concentrates

Twelve healthy blood donors participated on three (6 donors) or four (6 donors) occasions in the trial. 'A double bag apheresis set (Terumo, Japan), where the satellite bags were replaced with 300 ml P1-1240 bags (Travenol Sa, France), was used. The following procedure was repeated twice, giving two platelet concentrates from the same donor. 500ml whole blood was centrifuged immediately after donation at 2900 rpm for 90 seconds using a Damon PR-6000 centrifuge (International Equipment Company, USA). The red blood cells were reinfused and the platelet-rich plasma was expelled into the P1- 1240 bag and recentrifuged at 3400rpm for 8 minutes. The platelet-poor plasma was removed leaving approximately 70 ml plasma for platelet resuspension. The platelet concentrates were left undisturbed for 1 hour before agitation was commenced and they were thereafter agitated at room temperature using an obliquely-angled agitator (NOBEL Industries, Sweden). 25

Jaremo and Kutti Experimental protocol and filtration procedures

After 2 d of storage, two identical preparations were obtained by mixing and subsequently dividing these concentrates into two identical parts; thus, a total of 42 pairs of platelet concentrates were subject to study. One of the two preparations was filtered and the other served as the unfiltered control. The following filters were used: (a) Imugard IG500 (Terumo, Japan), (b) Miropore (Miramed, Italy), (c) Pall P1-100 (Pall, USA), and (d) Sepacell P1-1OA (Asahi, Japan). The filtration was performed according to the manufacturers' instructions. Consequently, the Pall P1-100 and Sepacell P1-1OA filters were used without pre-rinsing; the Imugard IG500 and Miropore filters were pre-rinsed with saline, and after the concentrates had passed freely into the filter they were again rinsed until the volume of the filtered concentrate was approximately 80 ml. Immediately after filtration, the platelet count, C3a des arg and the extracellular and total concentrations of PF4 and LDH were measured on the filtered as well as the unfiltered concentrates. On d 5 the extracellular concentrations of LDH and PF4 were determined in both concentrates.

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DONOR Fig. 1. Results for the individual donors (1 to 12) regarding the variations at the four (donor 1 to 6 ) and the three (donor 7 to 12) different donation occasions for the extracellular LDH levels in the unfiltered concentrates after 5 days of storage (* - not determined).

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Analytic procedures

The preparation of samples for the analyses of extracellular and total PF4 and LDH levels has been described previously (9). C3a des arg was determined on the supernatants obtained after centrifugation of the platelet concentrate at 15000 g for 5 min. Platelet counts were determined using a Coulter Counter S 8/80 (Coulter Electronics Ltd, UK), and LDH levels by standard procedures (10). RIA techniques were used for the determination of PF4 (Abbot, USA) and C3a des arg (Amersham, UK). The Wilcoxon non-parametric test for paired variables was used for the statistical evaluation. The results for Imugard IG500 and Miropore were corrected for the dilution during the filtration procedure. Results

Substantial variation as regards the extracellular LDH concentrations and the release of PF4 in the unfiltered concentrates were found on d 5 for the 12 individual donors at the different donation occasions (Figs. 1 and 2). All filters caused a significant (p < 0.01) decrease of the platelet counts (Table 1). Immediately after filtration the use of Imugard IG500 and Pall P1-100 decreased the extracellular PF4 activity significantly (p

Investigation of how different filters affect some biochemical properties of stored platelet concentrates.

The aim of the present study was to investigate how four different filters, i.e. Imugard IG500 (Terumo, Japan), Miropore (Miramed, Italy), Pall P1-100...
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