Coagulation Factor Activity in Platelet Concentrates T. L. SIMON A N D R. HENDERSON From the Puget Sound Blood Center. Seattle. Washington
Coagulation f8ctOr activity in platelet concentrates stored under diiferent conditions was Investigated. At both 22 C and 4 C, coagulationfactors 11, VII, IX, X, XI, and XI1 and fbrinogen were well maintained up to 72 hours. Factor V activity decllned slightly in platelet coneentratu stored at 4 C (to 78% at 72 hours), but fell to 47 per cent activity in platelets stored at 22 C. Factor VIII activity declined to approximately 68 per cent of normal activity by 72 hours at both 22 C and 4 C. Adtatlon did not make a difference in levels of txuphtlon factors. While the decline in Factor V was c k v l y more pronounced In platelet concentrates than in platelet-poor plasma from the same units, the decline in Factor VIII WM decreased by the presence of platelets. PIatekt coneentratu are a source of coagulation factor activity which should be considered in a component P-m.
BECAUSEPLATELET CONCENTRATES are transfused in bleeding disorders associated not only with thrombocytopenia but also with coagulation factor consumption, ( e . g . , leukemia, massive transfusion, cardiopulmonary bypass), an assessment of the coagulation factor activity transfused with such concentrates is important clinical information. The large volumes of plasma often transfused (e.g. 300-400 ml plasma in six units of 72-hour stored 22 C platelet concentrates) are a potential source of coagulation factor activity. Therefore, we have studied the level of coagulation factor activity in platelet concentrates stored for different intervals under different conditions. Methods Four hundred fitly ml whole blood was drawn from routine blood bank donors into Fenwal Supported by Blood Research and Demonstration Center grant HL-17265 from the National Heart, Lung and Blood Institute. Received for publication March 12, 1978; accepted April 25, 1978.
PL-146type blood bags, with 63 ml CPD anticoagulant. The platelet concentrates were prepared by the methods of Slichter and Harker.QsloTo separate platelet rich plasma from red blood cells, the blood was centrifugated at 1000 x g for nine minutes, at 22 C in a Sorval RC-3 centrifuge. The platelet rich plasma was centrifuged at 3000 x g for 20 minutes to obtain a platelet pellet which was then resuspended in the appropriate volume for 90 minutes at 22 C. Twenty to 30 ml volumes were used for 24 hour storage at 22 C; 20 to 35 ml volume for storage at 4 C for 72 hours, and 70 ml for storage at 22 C for 72 hours. Storage at 22 C conformed to the method of Slichter and Harker,'Oi.e. constant agitation at 22 2 C with concentrates in single layers on open wire racks mounted on a platform rotator (Eberbach, Ann Arbor), adjusted to 40 cycles per minute. Thirty-two units of platelet concentrate were stored for 72 hours at 22 C. At the end of the storage period, samples were obtained. Eight of the units were sampled at 24 and 48 hours for factor V and VIII levels. Eight platelet concentrate units were stored for 72 hours at 4 C. These were sampled at baseline, 24, 48, 72 hours for factors V and VIII and at 72 hours for all factors. Twelve additional platelet concentrate units were stored for 24 hours at 22 C and sampled at the end of the storage period for all the clotting factors. To assess the role of the platelet concentration in the plasma on factors V and VIII coagulation activity, seven units of platelet concentrate and platelet-poor plasma from the same donor were stored at 22 C for 72 hours under identical conditions. The effect of agitation and nonagitation of platelet concentrates was studied in four units of platelet concentrates stored and agitated at 22 C compared to four not agitated. Samples for coagulation factor activity were obtained by sterile technique from attached tubing after appropriate mixing. Samples were rendered platelet-poor by centrifugation at 30,000 x g. Coagulation assays were done by standard techniques. Normal (control) values for coagulation assays were determined by a minimum
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0041-1132/79/0300/0186$00.70 0 J. B. Lippincott Co. Tranafuaion Much-April 1979
186
Volume 19 Number 2
Volume 19 Number 2
187
COAGULATION FACTOR ACTIVITY
Table 1. Coaguletion Fector Activity in Platelet Concentrates under Different Storage Conditions Fibrinwen (mW)
Factor
Storage 22 c 72 h n . 22 c 24 h n . 4c 72 h n .
II
V
VII
Vlll
IX
X
XI
XI1
8 5 2 13
4 7 2 18
103t28
68222
96229
85221
96226
106t29
265
2
52
91 209
78221
117227
85+29
106225
9 3 t 16
1 0 6 2 19
107233
278
2
41
114224
7 8 2 10
8 9 * 16
6 7 2 18
1 0 2 2 16
98227
8 8 2 18
12Of20
269
f
32
pool of 10 ml blood from 20 different normal blood bank donors. Factor I1 was measured by comparing the ability of the test plasma and normal plasma to shorten the prolonged prothrombin time of substrate plasma. Factor I1 deficient plasma was made by mixing equal volumes of barium sulfate absorbed plasma and prothrombin-free serum.' Factors V, VII, and X were measured by a similar principal.l,ll Factor V deficient plasma was obtained from aged oxalated human p1asma.l' Factors VIII, IX, XI, and XI1 were measured by a one-stage assay comparing the abilities of test plasma and normal plasma to shorten the prolonged kaolin-activated partial thromboplastin time of the appropriate substrate Factor VIII and IX deficient substrate plasmas were obtained from patients with
Coagulation f8ctOr activity in platelet concentrates stored under diiferent conditions was Investigated. At both 22 C and 4 C, coagulationfactors 11, VII, IX, X, XI, and XI1 and fbrinogen were well maintained up to 72 hours. Factor V activity decllned slightly in platelet coneentratu stored at 4 C (to 78% at 72 hours), but fell to 47 per cent activity in platelets stored at 22 C. Factor VIII activity declined to approximately 68 per cent of normal activity by 72 hours at both 22 C and 4 C. Adtatlon did not make a difference in levels of txuphtlon factors. While the decline in Factor V was c k v l y more pronounced In platelet concentrates than in platelet-poor plasma from the same units, the decline in Factor VIII WM decreased by the presence of platelets. PIatekt coneentratu are a source of coagulation factor activity which should be considered in a component P-m.
BECAUSEPLATELET CONCENTRATES are transfused in bleeding disorders associated not only with thrombocytopenia but also with coagulation factor consumption, ( e . g . , leukemia, massive transfusion, cardiopulmonary bypass), an assessment of the coagulation factor activity transfused with such concentrates is important clinical information. The large volumes of plasma often transfused (e.g. 300-400 ml plasma in six units of 72-hour stored 22 C platelet concentrates) are a potential source of coagulation factor activity. Therefore, we have studied the level of coagulation factor activity in platelet concentrates stored for different intervals under different conditions. Methods Four hundred fitly ml whole blood was drawn from routine blood bank donors into Fenwal Supported by Blood Research and Demonstration Center grant HL-17265 from the National Heart, Lung and Blood Institute. Received for publication March 12, 1978; accepted April 25, 1978.
PL-146type blood bags, with 63 ml CPD anticoagulant. The platelet concentrates were prepared by the methods of Slichter and Harker.QsloTo separate platelet rich plasma from red blood cells, the blood was centrifugated at 1000 x g for nine minutes, at 22 C in a Sorval RC-3 centrifuge. The platelet rich plasma was centrifuged at 3000 x g for 20 minutes to obtain a platelet pellet which was then resuspended in the appropriate volume for 90 minutes at 22 C. Twenty to 30 ml volumes were used for 24 hour storage at 22 C; 20 to 35 ml volume for storage at 4 C for 72 hours, and 70 ml for storage at 22 C for 72 hours. Storage at 22 C conformed to the method of Slichter and Harker,'Oi.e. constant agitation at 22 2 C with concentrates in single layers on open wire racks mounted on a platform rotator (Eberbach, Ann Arbor), adjusted to 40 cycles per minute. Thirty-two units of platelet concentrate were stored for 72 hours at 22 C. At the end of the storage period, samples were obtained. Eight of the units were sampled at 24 and 48 hours for factor V and VIII levels. Eight platelet concentrate units were stored for 72 hours at 4 C. These were sampled at baseline, 24, 48, 72 hours for factors V and VIII and at 72 hours for all factors. Twelve additional platelet concentrate units were stored for 24 hours at 22 C and sampled at the end of the storage period for all the clotting factors. To assess the role of the platelet concentration in the plasma on factors V and VIII coagulation activity, seven units of platelet concentrate and platelet-poor plasma from the same donor were stored at 22 C for 72 hours under identical conditions. The effect of agitation and nonagitation of platelet concentrates was studied in four units of platelet concentrates stored and agitated at 22 C compared to four not agitated. Samples for coagulation factor activity were obtained by sterile technique from attached tubing after appropriate mixing. Samples were rendered platelet-poor by centrifugation at 30,000 x g. Coagulation assays were done by standard techniques. Normal (control) values for coagulation assays were determined by a minimum
*
0041-1132/79/0300/0186$00.70 0 J. B. Lippincott Co. Tranafuaion Much-April 1979
186
Volume 19 Number 2
Volume 19 Number 2
187
COAGULATION FACTOR ACTIVITY
Table 1. Coaguletion Fector Activity in Platelet Concentrates under Different Storage Conditions Fibrinwen (mW)
Factor
Storage 22 c 72 h n . 22 c 24 h n . 4c 72 h n .
II
V
VII
Vlll
IX
X
XI
XI1
8 5 2 13
4 7 2 18
103t28
68222
96229
85221
96226
106t29
265
2
52
91 209
78221
117227
85+29
106225
9 3 t 16
1 0 6 2 19
107233
278
2
41
114224
7 8 2 10
8 9 * 16
6 7 2 18
1 0 2 2 16
98227
8 8 2 18
12Of20
269
f
32
pool of 10 ml blood from 20 different normal blood bank donors. Factor I1 was measured by comparing the ability of the test plasma and normal plasma to shorten the prolonged prothrombin time of substrate plasma. Factor I1 deficient plasma was made by mixing equal volumes of barium sulfate absorbed plasma and prothrombin-free serum.' Factors V, VII, and X were measured by a similar principal.l,ll Factor V deficient plasma was obtained from aged oxalated human p1asma.l' Factors VIII, IX, XI, and XI1 were measured by a one-stage assay comparing the abilities of test plasma and normal plasma to shorten the prolonged kaolin-activated partial thromboplastin time of the appropriate substrate Factor VIII and IX deficient substrate plasmas were obtained from patients with
