Background: Red cell concentrates (RCC) would be required more in the aging society. Many blood donors usually donate whole blood once or twice a year. It would be useful if they could donate more blood in a single donation.
Objects and methods: Eighteen male donors with ≥58kg were enrolled and donated 3 units of RCC twice with a half year interval by automatic apheresis (RCa) machines (Haemonetics Co.). CCS was used in the first RCa (1-RCa) and Multi in the second RCa (2-RCa). These donors were asked the physical conditions and tested for blood cell counts, s-Fe, s-Ft, TIBC and free EPO during 6 months. The RCC collected was tested for blood volume, Hb, Ht, RBC, WBC, platelets, free Hb, ATP, and 2,3DPG.
Results: The donor's Hb and s-Ft values (mean±SD) before 1-RCa were 15.3±0.8g/dl and 87.2±50.6ng/ml, respectively. Slight citrate reactions were recognized by 5 donors during and feeling tired by other 3 after the procedures, but recovered without medication. The Hb values in both RCa did not decrease to <11g/dl soon after the procedures, and recovered to the previous levels at 3 months. The other laboratory findings except s-Ft changed almost the same in both RCa. The s-Ft values in 1-RCa and 2-RCa compared to their previous levels decreased to 45.8±31.8% (n=17) and 54.5±28.0% after 1month, but reached 61.8±20.2% (n=17) and 77.0±29.5% (n=13) at 6months, respectively. S-Ft levels with ≤12ng/ml were found in 6 donors within normal limits of Hb. Free Hb levels in the RCC collected by Multi were 20.1±10.8mg/dl soon after the procedures.
Conclusion: We consider that 3units of RCC could safely be collected twice a year with a half year interval by RCa, although repeated donations might result in continually lowering s-Ft levels.

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Background: For clinical cord blood (CB) transplantation, CB is processed using a standard hydroxyethyl starch protocol generally within 48 h of collection at room temperature. However, for tissue stem cell research, mononuclear cells (MNCs) were isolated from CB using a Ficoll-Paque density-gradient method. Here we report the effect of storage temperature and time from CB collection to processing on the cord blood mononuclear cells (CB-MNCs) isolated using a density-gradient method.
Methods: We processed CB using a Ficoll-Paque density-gradient method to collect the cells in the MNC layer. Cells were analyzed using an automatic blood cell counter, and CD34⁺ cells were counted according to the ISHAGE method.
Results: The recovery rate of viable MNCs in the CB-MNC layer was inversely related to the time from collection to processing of CB samples. However, recoveries of total nucleated cell and CD34⁺ were not affected by the time from collection to processing. The percentage of neutrophil contamination in the MNC layer increased significantly with increasing time from CB collection to processing (n=100, p<0.0001). Furthermore, CB stored at low temperatures had significantly less neutrophil contamination in the MNC layer than those stored at room temperature for 30 h after CB collection.
Discussion: Storage temperature and time from collection to processing influence the composition of CB-MNCs products processed using a Ficoll-Paque density-gradient method.

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Background: To prevent the risk of human parvovirus B19 (B19V) transmission through contaminated blood products, the Japanese Red Cross Blood Center has introduced B19V antigen screening by chemiluminescent enzyme immunoassay (CLEIA). Recently, B19V has been classified into three genotypes, 1, 2 and 3. According to the FDA guideline, B19V NAT for all plasma-derived products should detect all known genotypes of the virus. However, no survey for B19V genotyping or the possibility of detection of all genotypes by serology has been reported in Japan.
Methods: WHO panels including all genotypes of B19V were tested by universal real-time PCR (U-PCR) with some modifications and CLEIA to determine the sensitivity of each method. A total of 96 B19V DNA-positive donor samples obtained in Hokkaido during the past 13 years were assayed by U-PCR and phylogenetically analyzed by PCR direct sequencing.
Results: All genotypes of B19V were confirmed to be detected by U-PCR and CLEIA. The detection limits of U-PCR for genotypes 1, 2 and 3 were 13.6, 9.4 and 14.6IU/ml, respectively. The sensitivity of CLEIA was inferred to be approximately 6.3 log₁₀IU/ml. All 96 isolates from blood donors were detected by U-PCR and segregated into genotype 1.
Conclusion: All genotypes of B19V were detectable by CLEIA. B19V genotype 1 was the only strain circulating in Hokkaido during the past 13 years.

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Background: To prevent anti-D alloimmunization by D-positive RBCs derived from an infant or newborn, the administration of Rh human-immunoglobulin (RhIG) to D-negative women without anti-D antibody is recommended during pregnancy or at delivery. However, there is no guidance for pregnant woman with the weak D phenotype in Japan.
Case and Results: A primigravid woman was admitted to our hospital at 34 week's gestation. Her RhD typed as weak D because her RBCs were not agglutinated by the immediate spin method but were positive by the indirect antiglobulin test using a commercial anti-D reagent. RhIG was not administered to the woman during pregnancy or at delivery despite her first baby being D-positive. However, anti-D alloantibodies were not detected in her sera during pregnancy or at delivery. Her RHD gene was suspected to be D variant and analyzed. According to the sequencing results of the RHD gDNA and RHD cDNA derived from peripheral reticulocytes, the pregnant woman had a novel RHD gene with A>G mutation at the splicing acceptor site of intron 4. This intronic mutation may cause alternative splicing as a consequence of its generating a unique transcript with an 87 base insertion.
Conclusion: We experienced a case of a weak D primiparous woman in whom RhIG was not administered during the perinatal period or at delivery. Her first baby was D-positive. However, anti-D alloimmunization was not induced. She has a novel RHD(IVS4-2A>G) with A>G mutation at the splicing accepter site of intron 4. It was considered that abnormal splicing may have caused the weak D with a 29-amino acid insertion.

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Jr^a antigen is an extremely high-incidence antigen, with an incidence of the Jr (a-) phenotype of approximately 0.06% among Japanese. We report a Jr (a-) patient who was transfused with cross-match compatible random donor red blood cells (RBCs).
The patient was a male in his 50s, type A, RhD (+), who was admitted to our hospital for treatment of malignant lymphoma. As a previous blood donor, he was known to be Jr (a-). We confirmed that he did not have any irregular antibodies by antibody screening, and transfused him with random donor RBCs for anemia after chemotherapy. All RBCs transfused were found to be Jr (a+) on subsequent examination.
He received a total of 14 units from 8 donors, but no anti- Jr^a developed. This case suggests that Jr (a-) patients without anti-Jr^a may not need to be transfused with Jr (a-) RBCs.

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A questionnaire survey concerning laboratory tests done before transfusion was conducted in hospitals in Kanagawa Prefecture. In the 2007 fiscal year, 420 hospitals performed blood transfusions. Almost 70% of supplied blood units were used in 72 large hospitals with more than 300 beds. All tests, such as blood typing, antibody screening, and cross-matching, were done by medical technologists in these hospitals. Only 1.8% of red cell transfusions were done in small hospitals with under 19 beds, but the number of such hospitals is large (141/420; 33.6%). Of the 81 (57.4%) hospitals that answered the questionnaire, 64 (79.0%) did not employ medical technologists. Forty hospitals (40/64; 62.5%) consigned ABO typing and cross-matching to commercial clinical laboratories. Transfusion-related tests in the other 24 hospitals were done by doctors (21/24) or nurses (3/24). Both ABO blood typing and cross-match tests were done mainly using by low sensitivity methods, such as slide or saline tests (16/20). We have no reports about adverse transfusion reactions from these hospitals, but further study is necessary. Our meeting should discuss measures to insure the safety of transfusions at small hospitals without medical technologists.

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Recently, reduced-intensity cord blood transplantation (Mini-CBT) has been focused at many transplantation centers and it has been performed mainly for elderly patients. Although we find some reports that discuss transplantation results, there are still few papers that research transfusion medicine in CBT.
We have performed cord blood transplants to 11 patients with hematologically malignant disease. Five patients were conditioned by 12Gy of TBI (Total body irradiation) and high-dose chemotherapy (Full-CBT) and 6 patients were transplanted as Mini-CBT (2Gy of TBI, Fuludarabin and Busulfan). Post-transplant immunosuppressive regimen consists of cyclosporin+short term Methotrexate for Full-CBT and cyclosporin+mycophenolate mofetil for Mini-CBT. As post-transplant courses were uneventful in most cases and engrafted in all cases exhibiting less than mild GVHD, we could not observe significant differences in the transplantation results of both CBT groups. Regarding RBC transfusion in Full-CBT group, the frequency, total units, and final day of transfusion were 13.0, 26.0 and 43.0 respectively, and those in Mini-CBT group were 7.5, 15.0 and 36.5 respectively, resulting in no significant difference between the two groups. Regarding platelet transfusion, the frequency, total unit and final day of transfusion in Full-CBT group were 22.5, 217.5 and 43.5 respectively and those in Mini-CBT group were 23.0, 230.0 and 42.0 respectively, showing no significant differences. For trial, we used 5 unit-platelet concentrates in 2 cases, resulting in the large decrease of the amount of transfused platelets, but the frequency of transfusion showed no differences. No bleeding was observed in these cases.
As a conclusion Mini-CBT does not increase the quantity of consumed blood transfusions. However the number of case studies is still so small, further investigation is needed to reach final conclusions.

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The preparation unit of Hakodate Red Cross Blood Center (Hakodate BC) was consolidated with the Hokkaido Red Cross Blood Center (Sapporo BC) in 2006, and inventory of blood products is now controlled through a network system formed by blood centers in Hokkaido. Consolidation was projected to have a negative impact on the stable supply of blood products, especially platelet products, which have a short shelf life and thus a limited amount in inventory. We performed a retrospective investigation of the influence of consolidation, focusing on emergency orders of platelet products. Although platelet products supplied within two hours after an order were temporarily decreased immediately after consolidation in 2006, these recovered in 2009 to the level in 2005, a pre-consolidation year, by rearrangement of the inventory scale. Platelet products supplied within two hours after an order could be regarded as being derived from inventory stock. Platelet products supplied more than 6 hours after an order, which were not derived from inventory stock, were decreased in 2009 to half of that in 2006. Regarding platelet products that were ordered outside of business hours, 88.5% were supplied within one hour in 2009, in contrast to 53.8% in 2006. In conclusion, the stable supply of platelet products can be achieved by appropriate inventory management even in emergency situations.
Inventory management on a larger scale by the active inter-BC transport of blood products is thought to be an important way to achieve stable supply as well as economical efficiency, whereas management solely in a local blood center is difficult because of its small scale.

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Although blood transfusion is an essential and effective therapy, transfusion-related adverse events remain possible due to the allogeneic origin of blood products. In order to grasp and deal with these adverse events, a comprehensive reporting system, namely a hemovigilance system, needs to be established in Japan. We have been running a pilot study since 2007, in which all grades of transfusion-related adverse events are collected bimonthly using an online system. Seven university hospitals initially participated in this study, and were joined by five small hospitals (with fewer than 300 beds) in 2009. This paper reports the result of analysis of the data collected from 2007 to 2009.
Over the three years, the overall incidence of adverse events per bag was 1.50%. Platelet concentrate (PC) (4.34%) gave rise to statistically more adverse events (6-fold) than both red cell concentrate and fresh frozen plasma. This difference was possibly caused by allo-immunization of PC recipients, because these patients, who commonly have with hematological diseases, tend to receive regular blood transfusions, and thus to be frequently allo-immunized. In addition, we found that the incidence of adverse events varied between hospitals by number of beds and patient characteristics.
In conclusion, this online reporting system may be useful for the collection and analysis of actual adverse events in recipients, and may contribute to the development of a more comprehensive hemovigilance system in Japan.

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In Japan, about 4,000 hematopoietic stem cell transplantations (HSCT) are currently performed for various hematologic and non-hematologic disorders in about 200 hospitals per year. However, there have been no regulations or professional standards or guidelines for the processing of cellular therapy products routinely used for HSCT. Therefore, the Japan Society of Transfusion Medicine and Cell Therapy (JSTMCT), in collaboration with the Japan Society for Hematopoietic Cell Transplantation (JSHCT), have established guideline, titled 'the Japanese Standards for Processing Cellular Therapy Products Routinely Used for Hematopoietic Stem Cell Transplantation', for all hospitals and related personnel performing HSCT. According to a nation-wide survey performed by JSTMCT, it is likely that the number of medical staff and equipment is insufficient in many hospitals. Although these guidelines are based on the world-wide standard, the FACT-JACIE 3^rd edition, and are intended to be minimum standards, some modifications were made to reflect the present situation of most hospitals. The guidelines include; 1 Objective, 2 Application, 3 Product Collection, 4 Personnel, 5 Equipment and Facility, 6 Policies and Procedures, 7 Distribution, 8 Storage and Thawing, 9 Sample Storage, 10 Infusion, 11 Disposal, and 12 Provision. Appendices include outlines of each procedure related to transplantation and examples of standard operation procedures (SOPs) and record forms. The established standards are to be uploaded to the JSTMCT website so that individuals can access and download the SOPs and record forms, which can be revised for use at each hospital. An accreditation system is also planned to be established in the near future.

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[in Japanese]

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