1. 1 5/8/2015 Seven Day Storage At 4°C Of Previously -80°C Frozen AB Plasma Sm. Remco Strelitski, BSc Chief Lab technician, Military Blood Bank, Netherlands. NATO 2009

2. Borgman MA, et. al, J Trauma, Oct, 2007. TRANSFUSION AND TRAUMA Coagulopathy and massive transfusion Ratio 1:1 RBC:Plasma improves survival rates Early transfusion of plasma is recommended NATO 2009

3. 3 AB Plasma - 80°C ( 7 years) + 30°C 30 min AB FFP - 30°C ( ± 0.7 years) AB DFP Thaw Leukodepleted, apheresis unit of FFP (- 30°C) is thawed, repacked and frozen to - 80°C at the MBB Thaw Freeze In theater, the unit is thawed and warmed to + 30º C NLD MILITARY BLOOD BANK Production of -80°C frozen plasma

4. NLD MILITARY BLOOD BANK Products preparation time and availability 5/8/2015 4 Product-80ºC22ºC4ºCProduction Time RBC10 years 6 hours2 weeks90 min Platelets2 years6 hoursNot possible 45 min Plasma7 years6 hours?35 min AIM OF STUDY Improved availability of plasma: Storage time of thawed AB -80°C Plasma at 4°C? NATO 2009

5. Pilot: storage of units at 4°C for 6 weeks Storage of samples in vials at -80°C 5/8/2015 5 FVFVIIFVIII APTTPTFib Data of frozen samples were different from fresh samples Project continued with fresh samples NATO 2009

6. RESULTS APTT and Factor VIII 5/8/2015 6 NATO 2009

7. 5/8/2015 7 RESULTS INR (PT), Factor V and Factor VII NATO 2009

8. RESULTS 5/8/2015 8 Average product volume 310±7 mL Average fibrinogen content 0,95 gram/unit at day 7 NATO 2009

9. Clinical relevance of guidelines: 70% vs 50%? 5/8/2015 9 Coagulation time (sec) Activity (%) FVII FV FVIII NATO 2009

10. Conclusions At 7 days FVIII and FV activity are well above 50% Fibrinogen activity is reduced on day 14 at 4°C -80°C AB Plasma can be stored for 7 days at 4°C without clinically significant loss of its clotting capacity Since April 2009, 137 stored units have been transfused without any untoward effects reported In addition, mutual support between MTF in Afghanistan became easier by using 4°C cold chain. 5/8/2015 10 NATO 2009

11. 11 Platelets DMSO (5% w/v) - 80°C (2 years) + 30°C Platelets DMSO (5%, 14 ml) AB Plasma 5 min Platelets DMSO ( 0.2%) + AB Plasma + 30°C 30 min AB DFP - 80°C Leukodepleted unit of (apheresis) platelets is frozen and concentrated in 5% DMSO plasma. In theater, after 5 min of thawing, the PLT’s are reconstituted in thawed -80°C plasma Future application 4°C plasma Production of thawed -80°C frozen platelets?

12. 5/8/2015 12 Product-80ºC22ºC4ºCProduction Time RBC10 years 6 hours2 weeks90 min Platelets2 years6 hoursNot possible 10 min? Plasma7 years6 hours7 days35 min AIM OF STUDY Improved availability of plasma: Storage time of thawed AB -80°C Plasma at 4°C? NATO 2009 Future application 4°C plasma Production of thawed -80°C frozen platelets?