A Simple Method for Obtaining Platelet Concentrates Free of Aggregates

A Simple Method for Obtaining Platelet Concentrates Free of Aggregates A simple method is presented for obtaining platelet concentrates in a homogeneous suspension by centrifuging platelet-rich plasma at 25 C and allowing the centrifuged platelets to incubate a t this temperature for 30 minutes before resuspending them. Platelet concentrates (PC) prepared by the usual methods frequently contain aggregates which may cause technical difficulties during platelet transfusion. Aggregation also appears to be associated with platelet damage since the amount of certain platelet enzymes released into PC was reduced when aggregation was minimized by adding adenosine to, or reducing the pH of, the platelet-rich plasma (PKP) before centrifugation.1 T h e procedure t o be described provides PC free of aggregates without adding potentially contaminating substances that may have a deleterious effect o n other blood components. T h e method is a modification of the procedure reported by Pert and co-workers2 PKP is obtained by centrifuging one unit (about 500 ml) of whole blood containing acid citrate dextrose (ACD)a as anticoagulant i n a triple plastic bagb a t 2,500 x g" for 100 seconds a t 1OC. T h e PKP is expressed into the satellite bag and brought to 25 C by gently agitating it a t room Received for publication July 27, 1967; accepted October 12, 1967. Present Address: The Brocrklyn-cumberland Medical Center, Brooklyn, New York 11205. a NIH Formula A. b Fenwal Laboratories, Morton Grove. 111. c Sorval RC-3 Ccntrifugc with Swinging Bucket Head. Ivan Sorvall Inc., Norwalk, Conn. d Sorvall RC-3 Ckntrifuge with Angle Head. Ivan Sorvall Inc.. Norwalk, Conn. temperature for 10-15 minutes or in a 25 C water bath for 5 minutes. I t is then centrifuged a t 5,100 x gd for 10 minutes at 25 (:. After all b u t 10-15 ml of the platelet-poor plasma has been expressed into the third bag, the bag containing the platelets is allowed to stand a t room temperature for 30 minutes. T h e platelets are resuspended in the small volume of residual plasma by stroking the bag 10 times in 30 seconds by hand or by photographic roller. No aggregation was observed i n the 1: either ' C macro- or microscopically. Only about 2 per cent of the platelet enzymes nucleoside diphosphokinase, 3-phosphoglycerate kinase and enolase was released, as measured by the method previously described.1 I n contrast, when the temperature of the centrifuged platelets was 0 maintained a t 0 C for 1 minutes prior to resuspension, the PC contained many aggregates, and 10 to 20 per cent of these platelet enzymes was released. Also, aggregation and release of about 10 per cent of these enzymes were observed when the platelets were resuspended immediately after centrifugation a t 25 C. T h i s method is reproducible a n d provides platelets capable of supporting clot retraction. Large-scale in vivo testing will be necessary to evaluate the clinical efficacy of these preparations. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Transfusion Wiley

A Simple Method for Obtaining Platelet Concentrates Free of Aggregates

Transfusion, Volume 8 (1) – Jan 2, 1968

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Publisher
Wiley
Copyright
1968 AABB
ISSN
0041-1132
eISSN
1537-2995
D.O.I.
10.1111/j.1537-2995.1968.tb02389.x
Publisher site
See Article on Publisher Site

Abstract

A simple method is presented for obtaining platelet concentrates in a homogeneous suspension by centrifuging platelet-rich plasma at 25 C and allowing the centrifuged platelets to incubate a t this temperature for 30 minutes before resuspending them. Platelet concentrates (PC) prepared by the usual methods frequently contain aggregates which may cause technical difficulties during platelet transfusion. Aggregation also appears to be associated with platelet damage since the amount of certain platelet enzymes released into PC was reduced when aggregation was minimized by adding adenosine to, or reducing the pH of, the platelet-rich plasma (PKP) before centrifugation.1 T h e procedure t o be described provides PC free of aggregates without adding potentially contaminating substances that may have a deleterious effect o n other blood components. T h e method is a modification of the procedure reported by Pert and co-workers2 PKP is obtained by centrifuging one unit (about 500 ml) of whole blood containing acid citrate dextrose (ACD)a as anticoagulant i n a triple plastic bagb a t 2,500 x g" for 100 seconds a t 1OC. T h e PKP is expressed into the satellite bag and brought to 25 C by gently agitating it a t room Received for publication July 27, 1967; accepted October 12, 1967. Present Address: The Brocrklyn-cumberland Medical Center, Brooklyn, New York 11205. a NIH Formula A. b Fenwal Laboratories, Morton Grove. 111. c Sorval RC-3 Ccntrifugc with Swinging Bucket Head. Ivan Sorvall Inc., Norwalk, Conn. d Sorvall RC-3 Ckntrifuge with Angle Head. Ivan Sorvall Inc.. Norwalk, Conn. temperature for 10-15 minutes or in a 25 C water bath for 5 minutes. I t is then centrifuged a t 5,100 x gd for 10 minutes at 25 (:. After all b u t 10-15 ml of the platelet-poor plasma has been expressed into the third bag, the bag containing the platelets is allowed to stand a t room temperature for 30 minutes. T h e platelets are resuspended in the small volume of residual plasma by stroking the bag 10 times in 30 seconds by hand or by photographic roller. No aggregation was observed i n the 1: either ' C macro- or microscopically. Only about 2 per cent of the platelet enzymes nucleoside diphosphokinase, 3-phosphoglycerate kinase and enolase was released, as measured by the method previously described.1 I n contrast, when the temperature of the centrifuged platelets was 0 maintained a t 0 C for 1 minutes prior to resuspension, the PC contained many aggregates, and 10 to 20 per cent of these platelet enzymes was released. Also, aggregation and release of about 10 per cent of these enzymes were observed when the platelets were resuspended immediately after centrifugation a t 25 C. T h i s method is reproducible a n d provides platelets capable of supporting clot retraction. Large-scale in vivo testing will be necessary to evaluate the clinical efficacy of these preparations.

Journal

TransfusionWiley

Published: Jan 2, 1968

References

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