Plateletpheresis concentrates produced with the COMTEC cell separator: the French experience.

The latest generation of cell separators such as Trima (Gambro), Amicus (Baxter) and AS-TEC 204 (Fresenius), allow the collection of leucocyte-reduced platelet concentrates without secondary filtration. Fresenius has recently developed the COMTEC cell separator whose performance has been evaluated by several teams in France. This new cell separator is an improved version of the Fresenius AS-TEC 204 cell separator, designed to allow more efficient platelet collections. This study reports on the experience of six French teams (from Bordeaux, Clermont-Ferrand, Creteil, Dijon, Lille and Nancy) who obtained 696 leucocyte-reduced plateletpheresis concentrates in the course of collection using the new Fresenius COMTEC cell separator. All healthy volunteer donors fulfilled French selection criteria for platelet apheresis. Donors were eligible if they had suitable venous accesses, if their bodyweight was *50 kg and if their pre-apheresis platelet count was >150 x 10(9) l(-1). Between 4606 and 5229 ml of blood were processed. The mean volume of the platelet concentrates was between 439 and 493 ml (mean 460 +/- 63 ml). The platelet yield was of the order of 5.18 +/- 1.02 x 10(11) with only one platelet concentrate below the norm of 2 x 10(11) platelets (0.91 x 10(11)). No plausible explanation for this was found. The residual leucocyte levels conform to current norms. The platelet concentrates contained less than 1 x 10(6) leucocytes per concentrate (mean 0.233 +/- 0.150 x 10(6) leucocytes) in more than 97% of the components produced with >95% statistical confidence. The efficacy of the cell separator (52.44 +/- 7.35%) is comparable to that of other separators. The Fresenius COMTEC cell separator makes it possible to obtain leucocyte-reduced platelet concentrates which comply with current standards both in terms of platelet content and residual leucocyte level.

[Computerization of a bank of platelets with HLA phenotype and preserved at -196 degrees C]. / Informatisation d'une banque de plaquettes phénotypées HLA et conservées à -196 degrés C.

In order to complete the logistics of the unit cytapheresis, we have developed a decision helping system which rests upon the computer management of -196 degrees C cryopreserved HLA type platelets. The hardware used is a micro-computer equipped with two floppy disks and a printer. Two files have been created: namely "Product" and "Patient". Data relating to 800 platelet concentrates may be recorded on a floppy disk. The software which has been developed has several functions: 1 - The input of the parameters of a HLA type concentrate with the possibility of reservation for the given recipient; 2 - The print out of the bank's stock; 3 - The selection and the reservation of HLA matched platelets according to the recipient; 4 - The print out of concentrates allocated to each patient, it is possible to thaw out chosen platelet concentrate and a thawing report is printed out, thus enabling one to locate the concentrate in the bank. Thanks to this "aid in decision making", the management of - 196 degrees C cryopreserved HLA type platelets may be carried out. The proposed software allows an one line answer for any emergency transfusion case. Considering the recipient HLA typing the suitable best match platelet concentrates are instantly transfused or save.

[Preparation of leukocyte-depleted human platelet concentrates by centrifugation and filtration of a pool of sterilely connected buffy-coats]. / Préparation de concentrés de plaquettes humaines déleucocytés par centrifugation et filtration d'un pool de buffy-coats connectés stérilement.

We describe a new method for the preparation of standardised therapeutic doses of leukocyte depleted platelets. The first step is to remove the buffy-coat from whole blood units drawn on triple Siamese ACD/SAGM bags (Maco-Pharma) by means of a Compomat (NPBI). The second step is to connect (SCD Haemonetics) six buffy-coats and one plasma to a special kit (Maco-Pharma) including a PALL PL 100 filter; after centrifugation, the supernatant platelet concentrate is extracted, filtered and recovered in a 2 litre TOTM PVC bag. The volume, the number of platelets and leukocytes of these pools are measured. A comparison of these parameters is made with therapeutic doses prepared in the same way without filtration. Besides, pH measurements up to the 6th day of storage and bacteriological checks are carried out. The results show: no platelet loss related to filtration; a synergy between the preparation process out of buffy-coats and the filtration: so each dose contains less than 10(6) leukocytes; a good pH level allowing the storage for five days as it is associated to the bacteriological safety of the functionally closed system. This technique makes it possible to transfuse only leukocyte depleted platelet concentrates. In addition, it offers new prospects for standardisation and quality improvement.

Comparison between a solid-phase low-ionic-strength solution antiglobulin test and conventional low-ionic-strength antiglobulin test: assessment for the screening of antierythrocyte antibodies.

A solid-phase low-ionic strength salt antiglobulin test (LISS-SPAT) has been developed using a microplate coated with dried sera as a solid phase. Before coating, the in vitro C3d fragment generation was activated by adding heat-aggregated immunoglobulin. The LISS-SPAT was compared with low-ionic strength conventional antiglobulin test (LISS-AGT) and also with a test using polybrene or papain microplates. When detecting the IgG and IgM antierythrocyte antibodies the reaction was developed in the same way in LISS-SPAT and LISS-AGT. In routine work, the LISS-SPAT provides a fast, reliable, handy and inexpensive screening of antibodies. This method appears to be an additional method to the papain and polybrene tests in microplates.

[Assessment and value of the polybrene microplate test for study and identification of irregular anti-erythrocyte antibodies]. / Evaluation et intérêt du test au polybrène en microplaque pour la recherche et l'identification des anticorps irréguliers anti-érythrocytes.

We have adapted Lalezari's manual polybrene test for use with microplate technology for screening and identification of anti-erythrocytes antibodies with a view to future automation. The technical conditions have been standardized, firstly by using a programmable centrifuge and a sequential shaking, secondly by using a preservative medium for panel after dispensing onto microplates. This methodology has been run in parallel with papain test and LIS indirect antiglobulin test: 7,000 screenings have been performed and their results are considered here. Our results are comparable to those described for automatic and manual techniques. The polybrene-microplate test affords a fast, reliable, handy and inexpensive means of screening and identification for irregular antibodies. It appears as an additional method for enzymatic tests in microplate. An antiglobulin test can be carried out after negative tests.

[Viability of human red blood cells preserved for 35 days after leukocyte depletion (in vitro study)]. / Viabilité des globules rouges humains conservés pendant 35 jours après déplétion en leucocytes (étude in vitro).

24 leukocyte poor red cells concentrates (L.P.R.C.) were prepared by sterile connection of a leucocyte filter between the primary bag and the SAGM bag of a blood unit after centrifugation. Their quality was followed up to 42 days by means of a panel of tests including, ATP and 2,3-DPG levels, hemolysis, plasma potassium, lactate and glucose, and counts of the microaggregates. 24 standard units acted as a control group. Results showed better preservation of LPRC and especially less hemolysis, higher ATP levels and at least equal oxyphoric capacity (explored by 2,3-DPG). Microaggregate formation was dramatically reduced and bacteriologic checks (48 at day 25 and 48 at day 42) were all negative. Leucocyte depletion appears as a new way to improve functionality of erythrocytes during storage in the SAGM medium. 35 days shelf life will allow this blood product to be more available and its preparation more standardised.

[Leukocyte removal in a closed system of human red-cell concentrates: a technic coupled with the automated extraction of the buffy coat using a sterile connector from a filtration kit]. / Déleucocytation en système clos des concentrés de globules rouges humains: une technique associant l'extraction automatisée du buffy-coat à la connexion stérile d'un kit de filtration.

A technique, integrally run in a closed system, for leucocyte depletion of human red cell concentrates is described. It associates two complementary processes: buffy-coat removal and filtration. The first step is carried out with an automated system for blood component preparation (Compomat, NPBI); its efficiency is improved by a custom made blood collection set with ACD anticoagulant solution in the primary bag. The second step is simplified by a filtration kit requiring only one sterile connection for operation (SCD 312, Dupont de Nemours) and allowing a standardised rinsing of the filter. Quality control of 33 units so prepared shows principally: --an intensive leuko-depletion (4 logs) enabling leukocyte contamination to be kept below 10(6) per unit; --a moderate red cells loss (15 ml for the first step and 20 ml for the second one). This technique provides a permanently available and very pure blood component. Moreover it offers new potential for standardisation and mastery of quality control.