Vox Sanguinis (2015) 108, 160–168 © 2014 International Society of Blood Transfusion DOI: 10.1111/vox.12206
ORIGINAL PAPER
Thrombin generation and coagulation factor content of thawed plasma and platelet concentrates P. Cookson,1 A. Lawrie,2 L. Green,2 E. Dent, S. Proffitt,1 S. Bashir,1 S. Thomas1 & R. Cardigan1 1
Component Development Laboratory, NHS Blood & Transplant, Brentwood, UK Haemostasis Research Unit, Department of Haematology, University College London, London, UK
2
Background We assessed the haemostatic capacity of thawed plasma produced after ambient storage of whole blood for 24 h (RTFP24), and the supernatant of buffy-coat derived platelet concentrates (PC). Methods Platelet concentrates (n = 20) were tested on days 1, 5 and 7 of storage at 22°C and RTFP24 (n = 10) immediately following thawing and after 4 and 6 days storage at 4°C. Coagulation factor activity, thrombin generation – an activator of protein C (PROTAC) and rotational thromboelastometry (ROTEM) were assessed. Results In plasma and buffy-coat derived PC, there was a < 10% loss of factors II, IX and FX, but much higher loss of factors FV, FVII and FVIII. In plasma, the total or peak amount of thrombin generated was unaffected by storage for 6 days, with or without Protac, but there was an increase in lag time and decreased rate of clot formation by ROTEM. In PC, but not plasma, there was a 16% increase in FXII activity and increase in resistance to activated protein C, co-incidental to 30% loss of free protein S. Received: 22 April 2014, revised 20 August 2014, accepted 15 September 2014, published online 3 December 2014
Conclusions These data suggest thrombin generation is relatively unaltered when RTFP24 is thawed and stored for 6 days, and that the supernatant of PC has significant haemostatic capacity. Key words: blood components, blood processing, platelet concentrates plasma.
Introduction There are a variety of methods for producing plasma for transfusion, the guidelines for which differ internationally. In the United States, fresh-frozen plasma (FFP) must be frozen within 6–8 h of collection. Plasma produced from whole blood that has been stored for up to 24 h at 4°C prior to separation is labelled as a different component (FP24), but is used for the same clinical indications as FFP with the exception of specific replacement of FV and FVIII. FFP and FP24 must be used within 24 h of thawing, but at that point can be relabelled as ‘Thawed Plasma’ and stored at 4°C for a further 4 days, that is 5 days from initial thawing [1]. In Europe and elsewhere, FFP may be prepared from whole blood that has been Correspondence: Rebecca Cardigan, NHS Blood & Transplant, Long Road, Cambridge CB2 OPT, UK E-mail: [email protected]
160
stored at 20–24°C for 24 h prior to separation and this product may be labelled the same as FFP produced 070 IU/nl FVIII [2]. This probably reflects samples having been subjected to an additional freeze-thaw step compared to routine
Table 1 Haemostasis parameters in plasma following thawing and storage at 4°C and during storage of platelets at 22°C Plasma
Platelets
Parameter
Baseline
PT ratio APTT ratio Fibrinogen (Clauss, g/l) FII (U/ml) FV (U/ml) FVII (U/ml) FVIII (U/ml) FIX (U/ml) FX (U/ml) FXI (U/ml) FXII (U/ml) Antithrombin (U/ml) Free protein S (U/ml) Protein C (U/ml)
105 114 248 096 085 094 072 085 094 071 094 092 085 095
– – – – – – – – – – – – – –
011 010 040 014 023 027 019 011 022 016 027 014 019 006
Day 4 106 124 250 095 077 084 049 081 093 071 092 092 084 094
– – – – – – – – – – – – – –
Day 6 020 010*** 040 016 018* 037 013*** 010*** 025 016 027* 014 020 005
109 123 248 093 076 079 048 080 091 071 092 091 083 094
– – – – – – – – – – – – – –
021 010*** 041 015* 018** 038* 013*** 010*** 023 015 027* 014 020 006
P-value
Baseline
03435
ORIGINAL PAPER
Thrombin generation and coagulation factor content of thawed plasma and platelet concentrates P. Cookson,1 A. Lawrie,2 L. Green,2 E. Dent, S. Proffitt,1 S. Bashir,1 S. Thomas1 & R. Cardigan1 1
Component Development Laboratory, NHS Blood & Transplant, Brentwood, UK Haemostasis Research Unit, Department of Haematology, University College London, London, UK
2
Background We assessed the haemostatic capacity of thawed plasma produced after ambient storage of whole blood for 24 h (RTFP24), and the supernatant of buffy-coat derived platelet concentrates (PC). Methods Platelet concentrates (n = 20) were tested on days 1, 5 and 7 of storage at 22°C and RTFP24 (n = 10) immediately following thawing and after 4 and 6 days storage at 4°C. Coagulation factor activity, thrombin generation – an activator of protein C (PROTAC) and rotational thromboelastometry (ROTEM) were assessed. Results In plasma and buffy-coat derived PC, there was a < 10% loss of factors II, IX and FX, but much higher loss of factors FV, FVII and FVIII. In plasma, the total or peak amount of thrombin generated was unaffected by storage for 6 days, with or without Protac, but there was an increase in lag time and decreased rate of clot formation by ROTEM. In PC, but not plasma, there was a 16% increase in FXII activity and increase in resistance to activated protein C, co-incidental to 30% loss of free protein S. Received: 22 April 2014, revised 20 August 2014, accepted 15 September 2014, published online 3 December 2014
Conclusions These data suggest thrombin generation is relatively unaltered when RTFP24 is thawed and stored for 6 days, and that the supernatant of PC has significant haemostatic capacity. Key words: blood components, blood processing, platelet concentrates plasma.
Introduction There are a variety of methods for producing plasma for transfusion, the guidelines for which differ internationally. In the United States, fresh-frozen plasma (FFP) must be frozen within 6–8 h of collection. Plasma produced from whole blood that has been stored for up to 24 h at 4°C prior to separation is labelled as a different component (FP24), but is used for the same clinical indications as FFP with the exception of specific replacement of FV and FVIII. FFP and FP24 must be used within 24 h of thawing, but at that point can be relabelled as ‘Thawed Plasma’ and stored at 4°C for a further 4 days, that is 5 days from initial thawing [1]. In Europe and elsewhere, FFP may be prepared from whole blood that has been Correspondence: Rebecca Cardigan, NHS Blood & Transplant, Long Road, Cambridge CB2 OPT, UK E-mail: [email protected]
160
stored at 20–24°C for 24 h prior to separation and this product may be labelled the same as FFP produced 070 IU/nl FVIII [2]. This probably reflects samples having been subjected to an additional freeze-thaw step compared to routine
Table 1 Haemostasis parameters in plasma following thawing and storage at 4°C and during storage of platelets at 22°C Plasma
Platelets
Parameter
Baseline
PT ratio APTT ratio Fibrinogen (Clauss, g/l) FII (U/ml) FV (U/ml) FVII (U/ml) FVIII (U/ml) FIX (U/ml) FX (U/ml) FXI (U/ml) FXII (U/ml) Antithrombin (U/ml) Free protein S (U/ml) Protein C (U/ml)
105 114 248 096 085 094 072 085 094 071 094 092 085 095
– – – – – – – – – – – – – –
011 010 040 014 023 027 019 011 022 016 027 014 019 006
Day 4 106 124 250 095 077 084 049 081 093 071 092 092 084 094
– – – – – – – – – – – – – –
Day 6 020 010*** 040 016 018* 037 013*** 010*** 025 016 027* 014 020 005
109 123 248 093 076 079 048 080 091 071 092 091 083 094
– – – – – – – – – – – – – –
021 010*** 041 015* 018** 038* 013*** 010*** 023 015 027* 014 020 006
P-value
Baseline
03435
