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Transfusion of blood and blood components is one of the widely used medical procedures. The responsibility for provision of blood and blood components lies with Polish blood transfusion centers (CKiK) substantively supervised by the Institute of Hematology and Transfusion Medicine. Hospital based blood banks, hospital wards, immunohematology laboratories are supervised by CKiK. Every year approximately 600 thousand people donate blood, more than 75% of which are regular donors. The annual number of donations is maintained at a constant level of about 1.3 million. The aim of this position paper is to present the current status, achievements and advancement in collection, preparation and testing procedures and methods used to obtain the best possible quality blood components dedicated for clinical use. The aim is also to show the structure of blood transfusion service in Poland. Collaboration of all organizational units involved in transfusion medicine is crucial for providing high quality health care for patients. Polish transfusiologists and blood transfusion officers have largely contributed to transfusion science with numerous publications and significant research work. The implementation of novel methods and ongoing research positions Polish blood transfusion service among those of highly developed countries.

Quality assurance and safety of hematopoietic stem cells (HSC) with special emphasis on bacterial and fungal contamination is the prerequisite for any transplantation procedure. The aim was to determine the incidence rate of such contamination during processing of transplantation material with regard to HSC source: peripheral blood stem cell (PBSC), bone marrow (BM), or cord blood (CB). Analysis involved autologous and allogenic products dedicated for patients and comprised in all 4135 donations, including 112 BM (2.70%), 3787 PBSC (91.60%), and 236 CB (5.70%) processed in cell bank over the period 1996–2016. Aerobic and anaerobic contamination was determined.Analysis of the 20-year data revealed 42 contaminated products: 25 PBSC (0.66% of tested units) and 17 CB (7.20% of tested units). No microbial contamination of BM products was detected. Overall percentage of contaminated products was 1.01%, mostly with Staphylococcus epidermidis (61.36%). Bacterial contamination rate at cell bank is relatively low and processing in a closed system does not seem as crucial as might be expected. This is particularly true for BM components. Equally important are evaluation of donor’s medical status and condition of the puncture site for collection of source material. Implementation of appropriate sample collection procedures should help minimize the risk of false-positive results due to environmental contamination.

Background Until now, markers of hepatitis E virus (HEV) infection have not been studied in blood donors throughout Poland, and no acute case of HEV infection has been closely documented or confirmed by HEV RNA detection. The prevalence of HEV infection markers, including HEV RNA in Polish blood donors and virus genotypes was investigated. Study design and methods In total, 12,664 individual donations from 22 Polish blood transfusion centers were tested for HEV RNA by transcription-mediated amplification. In addition, 3079 first-time donors sampled throughout Poland also were screened for antibodies to HEV. HEV RNA and immunoglobulin M-positive donations were confirmed using real-time reverse transcription-polymerase chain reaction and Western blotting, respectively. Results Ten donors were identified as RNA initial reactive (one of 1266 donors), and six (one of 2109) were identified as repeat reactive and confirmed by real-time reverse transcription-polymerase chain reaction or seroconversion. Sequence analysis identified HEV Genotype 3c in one donor and Genotype 3i in two others. On average, 43.5% of donors were immunoglobulin G-positive. Immunoglobulin G seroprevalence ranged from 22.7% to 60.8% in group ages 18 to 27 years and 48 to 57 years, respectively and differed between administrative regions from 28.9% in Podlasie to 61.3% in Wielkopolska. Thirty-nine of the donors were immunoglobulin M-positive, and seven donors were IgM positive only (0.2%). Of 37 immunoglobulin M-reactive samples tested by Western blot, 24 (64.9%) were confirmed. Conclusions The current results indicate a high level of HEV endemicity throughout Poland compared with other countries. There is an urgent need to consider the protection of recipients of blood components against transfusion-transmitted HEV infection.

Background Cord blood (CB) is a valuable source of hematopoietic stem cells (HSCs). Extended storage of CB is possible provided that validated cryopreservation procedures are used. The study objective was to determine optimal methods of CB cryopreservation. Study design and methods In the study we 1) compared the effect of two-step cryopreservation and controlled-rate freezing method on the postthaw quality of CB (Study A) and 2) evaluated the postthaw quality of HSC fractions isolated from CB with various methods and frozen with controlled-rate freezing method (Study B). The same cryoprotectant mixture was used for 20 CB units (Study A) and 122 CB units (Study B). Results In Study A, 13.79 × 10(8) and 13.29 × 10(8) initial white blood cell (WBC) counts decreased to 6.38 × 10(8) and 6.02 × 10(8) after thaw for the two methods, respectively. The mononuclear cell (MNC) counts decreased from 5.90 × 10(8) to 3.71 × 10(8) and from 5.64 × 10(8) to 3.47 × 10(8) dependent on the method. MNC viability decreased from 99.0% to 97.4% for the former and from 98.5% to 97.2% for the latter method. The differences were insignificant. In Study B, postthaw WBC recovery in HSC fractions was 74.4% to 103.5%, MNC recovery 106.4% to 118.5%, CD34+ cell recovery 102.5% to 150.2%, and MNC viability 94.1% to 97.4%. Conclusion Neither the cryopreservation procedure nor the freezing of isolated HSCs affected product quality, which may indicate that various freezing methods can be used for cell banking provided the they follow recommendations of good manufacturing practice and Directive 2004/33/EC.

The activity of the Polish Blood Transfusion Service (BTS) is based on the Public Blood Transfusion Service Act sanctioned by the Polish Parliament. There are 21 regional Blood Transfusion Centers

H. W. Reesink, S. Panzer, C. A. Gonzalez, N. Lena, P. Muntaabski, S. Gimbatti, E. Wood, M. Lambermont, V. Deneys, D. Sondag, T. Alport, D. Towns, D. Devine, P. Turek, M.-K. Auvinen, T. Koski, C. K. Lin, C. K. Lee, W. C. Tsoi, E. Lawlor, G. Grazzini, V. Piccinini, L. Catalano, S. Pupella, H. Kato, S. Takamoto, H. Okazaki, I. Hamaguchi, J. C. Wiersum-Osselton, A. J. W. van Tilborgh, P. Y. Zijlker-Jansen, K. M. Mangundap, M. R. Schipperus, D. Dinesh, P. Flanagan, O. Flesland, C. T. Steinsvag, A. Espinosa, M. Letowska, A. Rosiek, J. Antoniewicz-Papis, E. Lachert, M. B. C. Koh, R. Alcantara, M. Corral Alonso & E. Muniz-Diaz

Abstract 2113 Poster Board II-90 The presence of white blood cells, particularly lymphocytes, in blood products has been shown to contribute to the development of a variety of adverse events including both donor anti-recipient and recipient anti-donor responses. Therefore leukoreduction of blood products or inactivation of leukocytes is necessary. We are currently using gamma-irradiation (Radiator Gammacell 3000 Elan) to inactivate leukocytes in blood components. This study shows the comparison of irradiation and PRT treatment (Mirasol*®) on lymphocyte survival and inactivation in non-leukoreduced platelet concentrates (PCs). PRT treatment is a pathogen reduction technology that targets nucleic acids after exposure to riboflavin and UV-light. We analyzed 7 untreated (C), 7 PRT treated (M) and 7 irradiated PCs (RD) in our study. Non-leukoreduced buffy coats (mean volume 65 ml) were obtained from the Regional Blood Center, Warsaw. PCs were prepared by pooling 15 buffy coats (ABO identical) suspended in 3 plasma units, and dividing the pool into 3 equal-weight units in bags made of the same material. Following addition of 35 ml of riboflavin solution, M units were illuminated. The same volume of saline solution, was added to the C group and RD units. All PCs were then stored at 22°C with agitation for 5 days. Samples were removed on days 1, 3 and 6 for analysis. The lymphocyte survival rate was determined by 7AAD (7-amino-actinomycin D staining of dead cells, Becton Dickinson) and their activation by anti-CD69-APC staining (Becton Dickinson). Samples were also stained with anti-CD45-PE antibodies to identify and gate on lymphocytes. Samples were analyzed on the Becton Dickinson Cytometer FACSCanto I. No increase in the number of dead cells was observed during 6 days of storage in the C group. After 3 days of storage however, in the M group the percentage of dead cells was significant higher than in C and RD groups (Student t-test, p=0.004 and p=0.03, respectively). After 6 days, the percentage of dead cells in the M samples was 72% vs. 30% following irradiation. The percentage of 7AAD-positive cells was significantly higher compare to C samples, both in M (p=0,001) and in RD samples (p=0,004). The percentage of dead lymphocytes was also observed to be statistically higher in M samples than in RD (p= 0,001). Analysis of lymphocyte activation was performed on live (7AAD-negative, CD45-positive) cells only. CD69 expression ranged between 20% and 40% in all tested samples (C, M, RD) during 6 days of storage. On days 1 and 3 of storage, Mirasol treatment significantly reduced lymphocyte activation as shown by the ratio (test/control) of %CD69-positive cells (p=0,004 and p=0,001, respectively) and mean fluorescence expression intensity of CD69. Interestingly, after 6 days of storage, RD samples showed significant higher lymphocyte activation then C and M samples (p=0,03 and p=0,02, respectively). In summary, a significant increase of dead lymphocytes after 6 days of storage was observed in PRT-treated PCs. This increase was two-fold higher than in gamma-irradiated PCs. At the same time a decrease in lymphocyte activation during 6 days of storage was observed in PRT-treated PCs. Overall, the use of PRT achieves better leukocyte inactivation than gamma-irradiation. Disclosures: No relevant conflicts of interest to declare.