Blood Bag System Research Articles

Obituary: Ruby Nelly Irene Pietersz (1942–2018)

Ruby Pietersz, MD, PhD, passed away on 1 March 2018 at the age of 75. Her legacy: whole-blood separation into components by the buffy coat method, applicable as a routine procedure, currently used in many blood centres around the world. Ruby was born in 1942 in Malang, Dutch East Indies (now, Indonesia). Between 1944 and 1945, she was, with her mother, interned in a camp in Malang during the Japanese occupation. Ruby and her family moved to The Netherlands in 1952. She was trained as a laboratory technician, worked in a hospital in Zwolle and subsequently moved to Amsterdam in 1966 to work at the laboratory of haematology and blood transfusion of the Free University of Amsterdam. In 1967, she started studying medicine, mainly in the evening times, and became a medical doctor in 1977. Next, she became a specialist in internal medicine in 1982 and joined the Red Cross Blood Bank in Amsterdam. In that time, Hans Loos and Peter Prins, chemical engineers of the Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, had preliminary results to separate whole blood in red cells, plasma and a buffy coat, containing 90% of the platelets and 70% of the white cells. From these buffy coats, platelet concentrates could be made. As part of her thesis, Ruby developed a validated method to produce leucocyte-poor platelet concentrates from five buffy coats, which could be stored for 7 days at room temperature. This method is now used in routine all over the world in many blood transfusion centres and blood banks and has increased the provision of platelet concentrates for patients enormously. Ruby further optimized the production of red cell and platelet concentrates by integrating leucocyte-removing filters in blood bag systems. Another very important development, initiated by Ruby, was the standardized cooling of whole blood from body temperature to room temperature, and subsequent overnight storage of whole blood at ambient temperature. This facilitated the uniform separation of whole blood that was collected in the afternoon and evening, during business hours the next morning, which was an enormous logistic improvement. Overnight hold also reduced the risk of bacterial contamination. With spiking experiments, Ruby proved that a 16- to 20-h hold of whole blood at room temperature, in combination with subsequent removal of the buffy coat and the remaining white cells by filtration, could abrogate the risk of contamination with Yersinia enterocolitica. The buffy coat method in combination with overnight room temperature storage of whole blood quickly found acceptance all over the world. Ruby was particularly proud that Canadian Blood Services in Canada moved to the buffy coat method after 2006. Her research continued after Blood Bank Amsterdam became part of the national Sanquin Blood Supply organization in 1998, when she became manager of Research, Development and Education. She found great enjoyment in sharing her knowledge. She travelled to many countries to assist blood processing facilities to implement the buffy coat method. She received numerous visitors from other blood centres all over the world to learn how the buffy coat procedure worked in routine conditions. She set up national training courses for blood bank staff which still continue to this day, and trained many physicians, including haematologists and transfusion specialists. Her work was her life fulfilment. However, Dutch laws are strict and she had to retire at the age of 65 in 2007. She found new enjoyment in having more time to care for her family and friends. After her retirement, she did continue to travel abroad to help blood banks to set up their procedures. In 2014, she had an important role in the European IPA project to strengthen the Blood Transfusion Organization in the former Republic of Macedonia. Her last publication [Pietersz et al., Int J Clin Transfus Med 2016;3:55] can be regarded as her testament; Good Manufacturing Procedures, SOPs, training, validation, and traceability all were regarded extremely important by Ruby. When she retired, she was knighted as Officer in the Order of Orange-Nassau for her many achievements for society. She was an honorary member of the Dutch Transfusion Society, and of the BEST Collaborative, of which she was a founding member. Early January 2018, we were informed that she had an incurable kidney tumour. Ruby had the following motto in her PhD thesis: ‘People without friends are poor. I did not know I was so rich’. Ruby wanted this motto to be used on her mourning card, to reflect the warmth of true friendships that she had had in her life. She will be missed by her family, friends and colleagues around the world, especially for the kind person that she was.

Eurobloodpack: a common European design for blood bag systems with integral leucodepletion filters

Three EBA specified blood bag configurations ('Eurobloodpack') are described which are capable of meeting >80% of its member's requirements. These include a 'top-and-top' and two 'bottom-and-top' packs enabling aseptic, pre-donation collection of up to 40 ml of samples, 427.5-522.5 ml of whole blood and the preparation of an extensive range of blood components. Features currently beyond the scope of ISO standardisation have been controlled including: anticoagulant and additive volumes; collection needle and sampling system; transfer tubing; cross-match line; base label; leucodepletion filter performance; compatibility of access ports and transfusion sets. Eurobloodpack has significant advantages for blood services and blood bag manufacturers.

Performance characteristics of a novel blood bag in‐line closure device and subsequent product quality assessment

BACKGROUNDIn high-volume processing environments, manual breakage of in-line closures can result in repetitive strain injury (RSI). Furthermore, these closures may be incorrectly opened causing shear-induced hemolysis. To overcome the variability of in-line closure use and minimize RSI, Fresenius Kabi developed a new in-line closure, the CompoFlow, with mechanical openers.STUDY DESIGN AND METHODSThe consistency of the performance of the CompoFlow closure device was assessed, as was its effect on component quality. A total of 188 RBC units using CompoFlow blood bag systems and 43 using the standard bag systems were produced using the buffy coat manufacturing method. Twenty-six CompoFlow platelet (PLT) concentrates and 10 control concentrates were prepared from pools of four buffy coats. RBCs were assessed on Days 1, 21, and 42 for cellular variables and hemolysis. PLTs were assessed on Days 1, 3, and 7 for morphology, CD62P expression, glucose, lactate, and pH. A total of 308 closures were excised after processing and the apertures were measured using digital image analysis.RESULTSThe use of the CompoFlow device significantly improved the mean extraction time with 0.46 ± 0.11 sec/mL for the CompoFlow units and 0.52 ± 0.13 sec/mL for the control units. The CompoFlow closures showed a highly reproducible aperture after opening (coefficient of variation, 15%) and the device always remained opened. PLT and RBC products showed acceptable storage variables with no differences between CompoFlow and control.CONCLUSIONSThe CompoFlow closure devices improved the level of process control and processing time of blood component production with no negative effects on product quality.

Blood processing

Blood processing

Comparison of two sterile connection devices and the effect of sterile connections on blood component quality

Sterile connection devices (SCDs) are used to connect pieces of polyvinylchloride tubing between blood bag systems. After observing a slight decrease in inner diameter of tubing welded with the CompoDock S2 SCD, the effect of welded tubing on storage characteristics of white blood cell (WBC)-reduced red blood cells (RBCs) and platelet (PLT) concentrates was studied. Welds were made with Terumo SCD (T-SCD) or CompoDock S2, and unwelded tubing served as reference. Three WBC-reduced RBC units or 3 PLT concentrates were pooled and divided to prevent donor-dependent differences. The units were transferred 10 times over (1) tubing with a T-SCD weld, (2) a CompoDock S2 weld, or (3) unwelded tubing. RBCs were stored for 42 days and free hemoglobin (Hb) was measured; PLT concentrates were stored for 8 days and CD62P expression was measured, as markers for blood component quality (n = 10 paired experiments). WBC-reduced RBC units had similar hemolysis at the end of storage: 0.47 +/- 0.28, 0.47 +/- 0.35, and 0.49 +/- 0.38 percent of total Hb, for tubing with a T-SCD weld, a CompoDock S2 weld, or no weld, respectively (not significant). CD62P expression of stored WBC-reduced PLT concentrates was not significantly different between the groups: 20.3 +/- 5.1, 19.8 +/- 5.1, and 22.3 +/- 9.8 percent for tubing with a T-SCD weld, a CompoDock S2 weld, or no weld, respectively. The quality of blood components, measured as RBC hemolysis and platelet CD62P expression, is not adversely affected by the presence of a sterile connection in the tubing, made by either the CompoDock S2 or the T-SCD.

The mantra of blood safety: time for a new tune?

The mantra of blood safety: time for a new tune?

Blood component fractionation: manual versus automatic procedures

Over the last few years, quality system requirements have been introduced for blood components. The necessary compliance with standard productions will have a considerable impact on Blood Banks. The introduction of automated methods is the most satisfactory means to meet these requirements for blood component preparation. A new device has been developed to automate the fractionation of blood into components. We evaluated the efficacy of this instrument as compared to manual methods. A total of 218 units of blood have been collected, into several different commercial blood bag systems (77 into standard quadruple bag systems, 141 into bag systems with integrated in line filters), and used to evaluate the universality of the instrument. Whole blood units were processed using the Top/Top system and the Compomat G4 (Fresenius HemoCare). A separate program protocol was developed for each kind of bag. Use of the Compomat G4 resulted in a statistically significant ( p<0.001) increase of the hemoglobin in filtered red cell concentrates (RCC) in comparison with the manual procedure, and a similar trend, even not statistically significant, has been observed for filtered RCC. Regardless of bag systems, we were able to observe a statistically significant increase of platelets in the platelet concentrates (PCs), when comparing automatic versus manual procedure. The automated procedure has been shown to be fast, and easy for the operators. This device reliably produces acceptable blood components, and has been shown adaptable to use with different blood bag systems.

Nonobese diabetic-severe combined immunodeficient mice transplantation of volume-reduced and thawed umbilical cord blood transplants following closed-system immunomagnetic cell selection.

Protocols for the expansion of human umbilical cord blood (UCB) progenitors begin with the selection of CD34+ cells from stored frozen and thawed units. Use of an immunomagnetic selection procedure within a closed blood bag system for volume-reduced UCB transplants was evaluated, and the influence of CD34 cell selection on in vivo engraftment potential was studied. Eleven thawed buffy coat-processed UCB units were processed within a standard blood bag with a washing solution. In six independent experiments, the same dosage of 2 x 104 CD34+ cells from paired selected and nonselected samples was transplanted into NOD-SCID mice. In two experiments, cells from the negative fraction were also transplanted. The purity of CD34+ cells after selection was correlated with the removal of supernatant after the first washing step and therefore with adequate removal of damaged or dead cells (r=0.86, p < 0.01). Mice transplanted with unselected UCB cells had more human cells within their marrow than animals transplanted with selected cells (8.6 +/- 5.9% selected group vs. 19.8 +/- 14.2% unselected group; p=0.04), whereas no engraftment could be observed transplanting cells from the two negative fractions. A higher percentage of human CD45+ cells in the unselected group were found to be positive for CD38, CD14, CD33, and CD19, indicating a higher potential for these unselected progenitors to differentiate into myeloid cells and B cells. Processing of volume-reduced and thawed UCB transplants within a closed-bag system before immunomagnetic CD34+ cell selection allows for the preparation of CD34+ cells of significant purity at technically useful cell recoveries. However, these experiments indicate a potential impairment of engraftment capacity for the CD34+ cell-enriched fraction.

In vitro characteristics of red blood cell concentrates prepared from under- and overcollected units of whole blood and from a paediatric blood bag system.

The problem of how to deal with red blood cell concentrates (RBCs) prepared from under- or overcollected units of whole blood (WB) and how to collect blood from underweight persons arises in the context of autologous predeposit. To determine the quality of RBCs stored in PAGGS-M additive solution prepared from under- and overcollected units of whole blood and of PAGGS-M RBCs prepared from a paediatric 250-mL top outlet blood bag system we measured blood picture, haemolysis, K+, pH, ATP and 2,3-DPG on days 0, 10, 20, 30, 40 and 49 of storage. The volume of WB collected ranged from 150 to 600 mL in 50-mL increments (4 units per volume). Haemolysis was under 0.8% on day 49 in all RBCs prepared from WB donations between 200 mL and 600 mL. However, the day 49 haemolysis level of standard RBCs prepared from 450 mL of WB (0.15 +/- 0.03%) was reached earlier in RBCs from under- and overcollected units of whole blood. 2,3-DPG levels decreased rapidly between days 10 and 20 in all RBCs studied. RBCs from 450-mL donations showed acceptable ATP maintenance after 49 days (70.4% of day 0 value), while all other RBC ATP levels were below 50% of the day 0 level on day 49. In vitro quality data of RBCs prepared from a 250-mL donation in the paediatric blood bag system after storage for about 25 days were comparable to those after 49 days of storage of standard RBCs. Our results suggest that it is feasible to transfuse PAGGS-M RBCs prepared from under- as well as overcollected units of WB in the autologous setting. However, we strongly recommend shortening the storage period of such RBCs to maintain the quality level of standard RBCs.

Impact of various red cell concentrate preparation methods on the efficiency of prestorage white cell filtration and on red cells during storage for 42 days.

White cell (WBC) reduction prior to storage of red cell (RBC) concentrates may reduce the incidence of HLA alloimmunization and may improve the quality of stored RBCs. An integrated WBC-reduction filter system was tested after various RBC preparation procedures (from whole blood), and the influence of filtration on RBCs during storage for 42 days was investigated. Four additive system RBC preparation protocols were used. Units prepared from conventional triple blood bags were held for 4 to 6 hours at 22 degrees C, and then the RBCs were separated via a hard spin and filtration, performed immediately (Group 1) or after 18 hours' storage at 4 degrees C (Group 2). Units prepared from a top-and-bottom collection system were held at 22 degrees C for 4 to 6 or 22 to 24 hours; the centrifuged RBCs were filtered immediately after preparation (Groups 3 and 4, respectively, by holding time). WBC reduction and filtration time were analyzed. The impact of WBC filtration on pH, hemolysis rate, hemoglobin content, ATP, potassium glucose, and lactate was investigated weekly during storage for 42 days. Filtration reduced the mean WBC count by 3 to 4 log10, to 0.19 +/- 0.25 x 10(6), regardless of the RBC preparation method. Mean filtration times differed significantly between the groups and were longest for Group 1. Besides hemolysis and pH values, which were greater in all filtered units, no major differences were found in filtered and unfiltered RBCs during the storage interval. The efficiency of prestorage WBC filtration of RBCs was unaffected by the preparation procedure. However, the filtration time for RBCs freshly prepared in the conventional triple blood bag system without buffy-coat depletion was unacceptable. No major metabolic differences between filtered and unfiltered RBCs during 42 days of storage were found.

Umbilical cord blood processing: volume reduction and recovery of CD34+ cells.

One of the main problems for the establishment of umbilical cord blood (UCB) banks is the storage space needed for the frozen samples. The aim of this study was to find a method of reducing the volume of UCB units without major losses of the haematopoietic progenitor-CD34+ cells. The UCB was collected into a triple blood bag system, in which the anti-coagulant had been previously adjusted. The blood bag was first centrifuged for red cell depletion followed by a second centrifugation for plasma reduction. At this point, the main bag containing the white blood cell (WBC) rich fraction "buffy coat' (BC) and the "waste bag' were sealed and detached. Haematologic cell counts and CD34+ cell quantification were done in whole blood and in the isolated fractions. The average volume of the 19 UCB samples processed was 103 ml. Separation by centrifugation led to a mean volume reduction of 56% with red cell depletion of 59%. The white blood cell recovery was of 72% with a significant CD34+ cell recovery of 87%. This seems a promising method for cord blood volume reduction and enrichment of CD34+ cells.

The Selection of Plastic Materials for Blood Bags

The procedures used in the preparation of blood components together with the processes used in the manufacture of multiple blood bag systems impose a unique combination of requirements that severely limits the selection of plastics. Plasticized PVC, the plastic used in the first blood bags introduced by Carl Walter over 40 years ago, remains the material of choice today. Blood bag material research has focused on two areas: (1) the development of containers with increased gas permeability for the storage of platelet concentrates; and (2) the reduction or elimination of plasticizer contamination of stored blood components. This research has led to the development of several second-generation containers that have improved the quality and extended the allowable storage period of platelet transfusion products. Plastics virtually free of extractives are available for the storage of platelets and plasma, but elimination of plasticizers from RBC products has not yet been achieved.

Blood component processing technique and plasma quality.

To maintain a closed system during the preparation of blood components, including the removal of buffy coat, many centers use a quadruple blood bag additive solution system which in this study has been reduced to a cheaper triple bag system. The buffy coat and plasma were after centrifugation transferred to the first satellite bag and, after a second spin, the plasma separated from the buffy coat was transferred to the second satellite bag and stored for a fortnight at 4 degrees C. This resulted in a statistically significant increase in platelet factor 4 and elastase activity levels. No significant changes were found in the levels of C1-esterase inhibitor and kallikrein inhibiting activity, thrombin-antithrombin complexes, soluble fibrin, fibrinopeptide A and spontaneous proteolytic activity. The changes observed must be regarded as clinically insignificant. The platelet count is low enough to meet the requirements for platelet poor plasma. Using this blood component separation technique, one can reduce the CPD/additive solution 4-pack blood bag system to a less expensive 3-pack blood bag system.

輸液関連システムと注射剤の配合変化

The safety of the injection therapy has to be secured including that of the injection itself as well as that of the equipment used for injection, i.e. i.v. fluid administration systems. The contamination of insoluble particulate matter ascribed to coating of silicone oil over disposable syringe is being solved by the development of syringe using fluoro-resin laminated rubber. In the case of blood administration set and blood bag system, there are some in which the solvent used for adhesion has been eluted. Further, the relationship between the quality of materials of administration set and the adsorption of drugs has become clear. As regards the plastic container for infusion, insoluble particulate matter and adsorption of drugs have become a problem in connection with the quality of materials used and yet the structure of plastic/rubber combined cap is related to the formation of core, thereby making the improvement. In the case of plastic bag, that made of polyethylene does not adsorb drugs so much and the formation of insoluble particulate matter is a little. For the measures of drug interaction of injection, i.v. fluid administration systems have been applied and the safe and effective method of administration of injection will be established along with the development of I.V. delivery system.

Preparation of leukocyte-poor platelet concentrates from buffy coats by a new type of quadruple bag

A new quadruple blood bag system for collection of platelet concentrates is described. The principle modification consists of a smaller (150 ml) buffy coat bag in conjunction with SAG-mannitol for red cell preservation. Platelet yield and function were comparable to conventional techniques, leukocyte contamination was lower (1.7 +/- 1.3 x 10(7) per unit). The system is recommended for specialized institutions with a high demand of blood components.

Extension of platelet concentrate storage.

Extension of the storage time of platelet concentrates in a satellite bag which is part of a new blood bag system was studied by reinfusing autologous 51Cr-labeled platelets into normal volunteers, and measuring postinfusion platelet counts and bleeding times in patients requiring platelet transfusions. This satellite bag, made of polyvinylchloride plasticized with a new agent, was found to protect platelet concentrates against fall of pH better than other containers studied. This protection was felt to be due to the greater gas permeability of the new plastic. Mean in vivo recovery and half-life (greater than 31% and 3.3 days, respectively) of autologous reinfused platelets were satisfactory following 5 days of storage. Following 7 days of storage, mean recovery was 41 percent and half-life was 2.8 days. Peripheral platelet count increments in patients following platelet transfusions with concentrates stored 4 to 7 days in the new plastic were comparable to increments following transfusion of platelets stored 2 to 3 days in the other plastics studied. Bleeding times shortened in three of four patients receiving platelet concentrates stored from 4 to 6 days in the new plastic. Platelet concentrates stored in the new bag at 20 to 24 degrees C with flat-bed or elliptical agitation could be transfused for up to 5 days following phlebotomy with acceptable clinical results. The new plastic container is promising for storage of platelet concentrates for up to 7 days. Due to the higher pH of 50-ml platelet concentrates stored in bags made with the new plastic, the concentrates were superior at any storage interval to those stored in bags made of the other plastics studied.

Cryoprecipitate and the plastic blood-bag system: provision of adequate replacement therapy for routine treatment of haemophilia.

In a survey of 210 young haemophiliacs living in South-east England it was recently shown that, by modern standards, under 20% received adequate treatment (Dormandy et al., 1967). The long-term management of the remainder fell short of what can and should be achieved even with the limited resources at present available to the hospital service. It was found, for example, that the potency of the frozen plasma issued at the time of the survey by the North London Blood Trans fusion Centre was often too low to be of any clinical benefit. This led to a comparative trial of plasma processed at the haemophilia centre of the Royal Free Hospital and at the trans fusion centre. At the same time a number of technical pro cedures were explored, particular attention being paid to the selection of those which could be widely adopted. After the report of Pool and Shannon (1965) cryoprecipitate rich in factor VIII was prepared ; 537 bags of this have been used in the trial since April 1966. In the course of the trial 239 infusions were given for 121 episodes of bleeding in patients with haemophilia. A further 23 infusions were given to patients with von Willebrand's disease. The results of the latter have been reported elsewhere (Bennett and Dormandy, 1966). A total of 1,304 pints (in 500 ml. units) of blood were used. The 914 pints which were collected by the plastic-bag method and processed at the haemophilia centre yielded 2,019 blood components, which were used separately. Supernatant plasma left after the separation of cryoprecipitate was given to three patients with Christmas disease (factor IX deficiency) and to one patient with plasma thromboplastin antecedent (factor XI) deficiency. Specific assays for factor IX and factor XI activity were carried out on these preparations.