Current status of red blood cell manufacturing in 3D culture and bioreactors

Owing to donor-related issues, blood shortages and transfusion-related adverse reactions have become global issues of grave concern. In vitro manufactured red blood cells (RBCs) are promising substitutes for blood donation. In the United Kingdom, a clinical trial for allogeneic mini transfusion of cultured RBCs derived from primary hematopoietic stem cells has recently begun. However, current production quantities are limited and need improved before clinical use. New methods to enhance manufacturing efficiencies have been explored, including different cell sources, bioreactors, and 3-dimensional (3D) materials; however, further research is required. In this review, we discuss various cell sources for blood cell production, recent advances in bioreactor manufacturing processes, and the clinical applications of cultured blood.

A Case of False-negative Malaria Rapid Diagnostic Test Induced by Treatment with Doxycycline

Rapid diagnostic tests (RDTs) for malaria using antibodies against pan-Plasmodium antigen lactate dehydrogenase (pLDH) are commonly used for malaria diagnosis. The level of malaria parasitemia determined by peripheral blood smears (PBS) correlates with the pLDH concentration in most cases. We report a case of malaria recurrence associated with false-negative RDT results. A 22-year-old male patient was admitted to the Armed Forces Capital Hospital with fever and chills, and was diagnosed with malaria infection. Four days after antimalarial treatment, these symptoms recurred. After admitting to our hospital, doxycycline was administered for 4 days. Even after administration of doxycycline, the malaria parasites in blood smears remained positive, but RDT showed negative results. Therefore, for patients receiving doxycycline, serial blood smear testing should be performed to exclude false-negative malaria RDT results.

Reduction of the Platelet Transfusion Dose and Its Effects / 대한진단검사의학회지

BACKGROUND: In Korea, a platelet transfusion dose (TD) of 8 units of platelet concentrates (PC) is usually used. To minimize the shortage of blood products and transfusion-related adverse reactions, the TD has been changed from 8 to 6 units in 2006 in our hospital. Here, we analyzed the dose reduction effect on patients' platelet counts and transfusion frequency. METHODS: We compared the amount of issued PC, platelet counts before and after transfusion, post-transfusion platelet increments, and transfusion frequencies in patients who were transfused with 8 PC in 2006 and 6 PC in 2008. RESULTS: Despite an increase in the number of admitted patients by 20% in 2008 with a disease distribution similar to that in 2006, the number of issued PC in 2008 was decreased by 26.6% compared to that in 2006. In 2008, post-transfusion platelet counts, pre-transfusion platelet counts in patients transfused with 320 mL whole blood-derived PC, and platelet increments in patients transfused with 400 mL whole blood-derived PC were significantly decreased. However, the mean transfusion frequency per one month was not significantly different, 4.3 times in 2006 and 4.7 in 2008. CONCLUSIONS: By implementing a policy of platelet TD restriction, the amount of total issued PC was markedly decreased. Although post-transfusion platelet counts were decreased, the transfusion frequency in a month was not significantly increased. The restriction of platelet TD was helpful for increasing physicians' recognition of blood shortage while achieving similar transfusion effects. We conclude that 6 units of PC would be a better guideline for the platelet TD.

A Case of Immune Hemolytic Anemia Induced by Ceftizoxime and Cefobactam (Sulbactam/Cefoperazone) / 대한진단검사의학회지

Simultaneous drug-induced immune hemolytic anemia (DIIHA) caused by multiple drugs is rare. We report a case of a patient who developed DIIHA caused by 2 drugs. The patient's serum exhibited agglutination of ceftizoxime- or sulbactam-coated red blood cells (RBCs; via a drug-adsorption mechanism) and of uncoated RBCs in the presence of sulbactam (via an immune-complex mechanism). Although ceftizoxime is known to exhibit a positive reaction by an immune-complex method with or without reactivity with drug-coated RBCs, this patient's antibodies were reactive only against drug-coated RBCs. On the other hand, sulbactam, which is known to cause hemolytic anemia by nonimmunologic protein adsorption, exhibited positive reactions in tests with both drug-coated RBCs and in the presence of sulbactam. This is the first report of DIIHA due to a sulbactam-cefoperazone combination and the fourth report of DIIHA due to ceftizoxime. Owing to the patient's complicated laboratory results, DIIHA was suspected only at a late stage. We propose that for the prompt diagnosis of DIIHA, tests for all possible causative drugs should be conducted by 2 methods.

Analysis of Characteristics of Mononuclear Cells Remaining in the Leukoreduction System Chamber of Trima Accel(R) and Their Differentiation Into Dendritic Cells / 대한진단검사의학회지

BACKGROUND: We investigated the characteristics of the mononuclear cells remaining in the leukoreduction system (LRS) chambers of Trima Accel(R) in comparison with those of standard buffy coat cells, and evaluated their potential for differentiation into dendritic cells. METHODS: Twenty-six LRS chambers of Trima Accel(R) were collected after platelet pheresis from healthy adults. Flow cytometric analysis for T, B, NK, and CD14+ cells was performed and the number of CD34+ cells was counted. Differentiation and maturation into dendritic cells were induced using CD14+ cells seperated via Magnetic cell sorting (MACS(R)) Seperation (Miltenyi Biotec Inc., USA). RESULTS: Total white blood cell (WBC) count in LRS chambers was 10.8x108 (range 7.7-18.0x108). The median values (range) of proportions of each cells were CD4+ T cell 29.6% (18.7-37.6), CD8+ T cell 27.7% (19.2-40.0), B cell 5.5% (2.2-12.1), NK cell 15.7% (13.7-19.9), and CD14+ cells 12.4% (8.6-32.3) respectively. Although total WBC count was significantly higher in the buffy coat (whole blood of 400 mL) than the LRS chambers, the numbers of lymphocytes and monocytes were not statistically different. The numbers of B cells and CD4+ cells were significantly higher in the buffy coat than the LRS chambers (P<0.05). The median value (range) of CD34+ cells obtained from the LRS chambers was 0.9x10(6) (0.2-2.6x10(6)). After 7 days of cytokine-supplemented culture, the CD14+ cells were successfully differentiated into dendritic cells. CONCLUSIONS: The mononuclear cells in LRS chambers of Trima Accel(R) are an excellent alternative source of viable and functional human blood cells, which can be used for research purposes.

A Nationwide Survey Report on the Blood Transfusion Services in Korea / 대한수혈학회지

BACKGROUND: Inspecting thestatus of blood banks has been done for the larger Korean hospitals, but it has never been done for the smaller ones on a nationwide scale in Korea. Here, we analyzed the status of the blood banks for their transfusion services and equipment, and especially for the smaller hospitals. METHODS: The subjects were all the hospitals that were provided more than one unit of blood by the Korea Red Cross (KRC) in 2006. We divided the hospitals to a big-hospital group and a small-hospital group that received over or under 5,000 units of blood, respectively, from the Korea Red Cross in 2006. The questionnaires were delivered by mail. RESULTS: The number of total hospitals was 2,488 and the number of hospitals in the small-hospital group was 2,381, and this accounted for 95.7% of the total hospitals. The response rate was 23.1%. Among the small-hospital group, 35% had no working manual, 61% were not involved in certification programs and 17% had no refrigerators that were exclusively used for blood. Furthermore, 31% performed only cell typing as ABO typing, 69% didn't test for antibody detection, and 7% used a slide method for crossmatching tests. Only 6% used a blood information sharing system and only 28.4% of the hospitals shipped blood by blood transport containers. The mean amount of discarded blood was 16.8 units and the main component was RBC. CONCLUSION: The level of management and services showed a great difference between the two groups of Korean hospitals. The small-hospital group is thought to need more support and attention from the government. This study will supply essential data for understanding the current state of blood transfusion services and establishing government policies for safe transfusion.

A Rare Case of Transfusion-related Anti-K Alloantibodies / 대한수혈학회지

No abstract available.

The Trends for Nationwide Blood Collection and the Supply of Blood in Korea during 2002~2006 / 대한수혈학회지

BACKGROUND: The recent trends for blood collection and the blood supply were analyzed. METHODS: Data from the annual reports of the Korean Red Cross from 2002 to 2006 were analyzed. RESULTS: The number of donors in 2002~2003 was about 2,530,000, but this decreased to 2,300,000 in the past 3 years with the population's donation rate being 4.7%. By age, those donors between 16~29 years made up 83% of all the donors. As donor verification became possible in real-time, blood collection from the registered deferral donors was decreased. Blood drawn by the KRC made up 98% of all the blood collected in Korea. Plasma collection for fractionation had recently decreased because of the blood shortage for transfusion in hospitals. The collection of single donor platelets has increased to up to 25% of all the platelets used in Korea. The supply of pre-storage leuko-reduced RBCs had increased. The inventory levels of blood components were lower than the proper levels for most of the days in 2006. The rate of discarding outdated blood components was markedly decreased due to a shortage of blood. The positive rate in screening tests for transfusion-related infection was an average of 2.4%. By nucleic acid tests,which were initiated from 2005, 14 cases during the window period (10 cases of HCV and 4 cases of HIV) were detected. CONCLUSION: For insuring a safe supply of blood, the donor information systems and up-to-date tests were deemed to become of good quality. However, the blood shortage should be resolved as soon as possible to maintain a consistent blood supply.

The Trends for Nationwide Blood Collection and the Supply of Blood in Korea during 2002~2006 / 대한수혈학회지

BACKGROUND: The recent trends for blood collection and the blood supply were analyzed. METHODS: Data from the annual reports of the Korean Red Cross from 2002 to 2006 were analyzed. RESULTS: The number of donors in 2002~2003 was about 2,530,000, but this decreased to 2,300,000 in the past 3 years with the population's donation rate being 4.7%. By age, those donors between 16~29 years made up 83% of all the donors. As donor verification became possible in real-time, blood collection from the registered deferral donors was decreased. Blood drawn by the KRC made up 98% of all the blood collected in Korea. Plasma collection for fractionation had recently decreased because of the blood shortage for transfusion in hospitals. The collection of single donor platelets has increased to up to 25% of all the platelets used in Korea. The supply of pre-storage leuko-reduced RBCs had increased. The inventory levels of blood components were lower than the proper levels for most of the days in 2006. The rate of discarding outdated blood components was markedly decreased due to a shortage of blood. The positive rate in screening tests for transfusion-related infection was an average of 2.4%. By nucleic acid tests,which were initiated from 2005, 14 cases during the window period (10 cases of HCV and 4 cases of HIV) were detected. CONCLUSION: For insuring a safe supply of blood, the donor information systems and up-to-date tests were deemed to become of good quality. However, the blood shortage should be resolved as soon as possible to maintain a consistent blood supply.

Automated Collection of Leukoreduced Double Red Blood Cell Units using an Alyx Apheresis Device / 대한수혈학회지

BACKGROUND: Automated collection of leukoreduced double red blood cell (RBC) units has been used to improve blood collection efficiency in many countries. The aim of this study was to evaluate leukoreduced double RBC unit collection using apheresis and to determine its clinical uses. METHODS: Automated leukoreduced double RBC units were collected from 17 healthy male volunteers using an Alyx apheresis device (Baxter, DeerWeld, IL). Our institutional criteria for male donors were as follows: height > or =170 cm, weight > or =70 kg, and hemoglobin > or =14.5 g/dL. Each donor's complete blood count (CBC) was determined before and after the apheresis procedure. In order to validate the final leukoreduced double RBC units, white blood cell (WBC) counts were measured manually using a Nageotte chamber. RESULTS: Leukoreduced double RBC units were collected in an average of 27+/-7 min, without specific donor reactions. The average volume of one unit of leukoreduced RBCs was 281+/-6 mL. Each donor's hemoglobin (P<0.001) and hematocrit (P<0.001) were significantly lower after donation, though WBC and platelet counts were no different. No residual WBCs were found in the leukoreduced, filtered product upon Nageotte chamber counting. Twenty-eight units of leukoreduced RBCs from 14 donors were transfused to patients who required leukoreduced RBCs, without specific transfusion reactions. CONCLUSION: The Alyx apheresis device allowed for safe and effective collection of leukoreduced double RBC units from a single donor. Automatically collected leukoreduced double RBC units were transfused to patients for the first time in Korea.

Automated Collection of Leukoreduced Double Red Blood Cell Units using an Alyx Apheresis Device / 대한수혈학회지

BACKGROUND: Automated collection of leukoreduced double red blood cell (RBC) units has been used to improve blood collection efficiency in many countries. The aim of this study was to evaluate leukoreduced double RBC unit collection using apheresis and to determine its clinical uses. METHODS: Automated leukoreduced double RBC units were collected from 17 healthy male volunteers using an Alyx apheresis device (Baxter, DeerWeld, IL). Our institutional criteria for male donors were as follows: height > or =170 cm, weight > or =70 kg, and hemoglobin > or =14.5 g/dL. Each donor's complete blood count (CBC) was determined before and after the apheresis procedure. In order to validate the final leukoreduced double RBC units, white blood cell (WBC) counts were measured manually using a Nageotte chamber. RESULTS: Leukoreduced double RBC units were collected in an average of 27+/-7 min, without specific donor reactions. The average volume of one unit of leukoreduced RBCs was 281+/-6 mL. Each donor's hemoglobin (P<0.001) and hematocrit (P<0.001) were significantly lower after donation, though WBC and platelet counts were no different. No residual WBCs were found in the leukoreduced, filtered product upon Nageotte chamber counting. Twenty-eight units of leukoreduced RBCs from 14 donors were transfused to patients who required leukoreduced RBCs, without specific transfusion reactions. CONCLUSION: The Alyx apheresis device allowed for safe and effective collection of leukoreduced double RBC units from a single donor. Automatically collected leukoreduced double RBC units were transfused to patients for the first time in Korea.

A Case of Hemolytic Transfusion Reaction in a Patient with Anti-E, Anti-M, Anti-Jkb, and Anti-Lea / 대한수혈학회지

We reported a case of hemolytic transfusion reaction that was related to multiple RBC antibodies such as anti-E, anti-M, anti-Jkb and anti-Lea after serial RBC transfusions. A forty-nine year old female visited the emergency room (ER) with hematochezia. She had previously received 16 units of packed RBCs from 2003 to Jan 2007 for her intermittent esophageal varix bleeding. No specific antibodies were identified before this visiting. At the ER, under the request for packed RBCs, we identified anti-E antibody within her serum. Her blood type was AB, RhD+ with the phenotype of CcDe. She received 5 units of E antigen negative RBCs. However, she showed hemolytic transfusion reactions such as mild fever with a decrease of hemoglobin from 11.4 g/dL to 6.8 g/dL after the transfusion. From the 8th to the 10th hospital day, another 3 units of E-antigen negative with the least incompatible RBCs were transfused to the patient, but the level of hemoglobin was not definitely increased. At the 14th hospital day, she received a final 2 units of leuko-reduced RBCs without E, M and Jkb antigens. Her hemoglobin was increased right after the final transfusion. We found that the patient's serum reacted with multiple RBC antibodies such as anti-E, anti-M, anti-Jkb and anti-Lea antibodies. She finally recovered from acute varix bleeding and was discharged on the 26th hospital day with the level of hemoglobin being 8.3 g/dL.

High Dose Chemotherapy and Autologous Peripheral Blood Stem Cell Transplantation in Pediatric Patients with High-risk Neuroblastoma / 대한수혈학회지

BACKGROUND: High dose chemotherapy followed by autologous peripheral blood stem cell transplantation (PBSCT) has become standard therapy for high-risk neuroblastoma patients. We performed a retrospective analysis to assess the characteristics of peripheral blood stem cell harvest (PBSCH) and PBSCT and its clinical outcome. METHODS: We reviewed 17 cases of patients with high-risk neuroblastoma that underwent PBSCH and/or high dose chemotherapy followed by PBSCT. RESULTS: Sixteen patients had stage IV neuroblastoma and one patient had a stage III neuroblastoma with MYCN amplification. The median age of the 17 patients was 43 months (range 22~114 months) and the median body weight was 15 kg (range 10~24 kg). After induction chemotherapy using a modified N7 protocol, 34 PBSCHs (1.5 leukapheresis per PBSCH) were performed. A statistically significant correlation was found between the pre-leukapheresis CD34+ cell count and the total number CD34+ cells of the harvested products (P<0.0001). Tyrosine hydroxylase mRNA was not detected by RT-PCR in all of the leukapheresis products. High dose chemotherapy followed by PBSCT was performed in 24 cases. The mean infused CD34+ cell dose was 4.01x106/kg and WBC and platelet engraftment was performed on day 12.0 and 21.5, respectively. Eleven patients died, and six patients are surviving 11 to 68 months after PBSCT (median survival time, 35 months). CONCLUSION: In this single institution study, treatment with high dose chemotherapy and PBSCT was performed successfully for children with high-risk neuroblastoma.

Evaluating the Appropriateness of a Single Unit Transfusion / 대한수혈학회지

BACKGROUND: The domestic quantity of blood components consumed has been decreasing since 2002, but the rate of a single unit RBC transfusion (SUT) has been on the increase. In the past, a SUT was regarded as an uncesssary procedure, but currently is considered as an effective method to maintain a minimal hemoglobin concentration for physiological needs. We investigated the actual conditions of a SUT. METHODS: We analyzed 800 cases of SUTs performed at a tertiary care university hospital between March 2006 March and February 2007. The subjects of the study were divided into a surgical group (n=561) and medical group (n=239) for the purpose of RBC unit usage and were analyzed by groups and ordering departments, with an analysis of the pre and post-transfusion hemoglobin concentration and hematocrit values. The distribution according to the pre and post-transfusion hemoglobin ranges were calculated. RESULTS: The mean hemoglobin concentration increment of the surgical group was significantly lower than that of the medical group (P<0.0001) and the mean pre and post-transfusion hemoglobin concentrations of the medical group were lower than that of the surgical group (P<0.0001). Approximately 26% cases of the SUTs performed in the surgical group were appropriate, based on a post-transfusion hemoglobin concentration below 10 g/dL. In the medical group, about 75% of the SUTs were appropriate based on a pre-transfusion hemoglobin concentration below 9 g/dL. CONCLUSION: Most transfusions are decided based on various clinical situations and opinions of the clinicians. Therefore, continuous evaluation of the appropriateness of transfusion is necessary. In our study, the appropriateness of a SUT was estimated indirectly based on the pre and post-transfusion hemoglobin concentration. Consequently, policies and strategies for performing asingle unit RBC transfusion are required.

A Case of ABO Discrepancy due to Anti-IH / 대한수혈학회지

The patient was a 70-year-old woman with hypertension and end stage renal disease, and she presented with left wrist pain due to falling a day before admission. On admission, laboratory testing revealed a hemoglobin level of 6.7 g/dL, and a physician ordered 2 units of packed RBCs. She had never received a RBC transfusion in the past. The ABO grouping showed a discrepancy between the cell type AB and serum type O, and the irregular antibody screening was negative. Crossmatchings with group AB and group O RBCs were incompatible. Anti-I, which is a cold antibody, was inferred because the degree of agglutination was decreased after warming. However, crossmatching with group O RBCs, which are the universal donor blood, was positive and the anti-IH was considered to be the specificity of the irregular antibody. The patient's serum did not react with group O cord (i) blood cells and anti-I was then considered. The genotype of this patient was AB, and it was inferred that the ABO discrepancy was due to anti-IH.

A Reestablished Reference Interval of Plasma Anion Gap Minimizing Preanalytical Errors / 대한진단검사의학회지

BACKGROUND: The reference interval of anion gap established in the 1970s has been changing as the method of electrolyte measurement changes. It is also influenced by preanalytical errors. But there are only a few reliable reports that examined the effect of various preanalytical errors. Furthermore, there has been no report of values measured by the instrument being used in our laboratory. Therefore, we attempted to establish a reference interval of anion gap measured by Hitachi 7170s minimizing preanalytical errors, and analyzed the effect of a delay in electrolyte measurement and of sample exposure to the air. METHODS: The subjects were 538 healthy people who attended Hanyang university hospital health clinic with normal blood levels for albumin, creatinine and glucose. The plasma Na+, K+, Cl-, and total carbon dioxide (TCO2) were measured by Hitachi 7170s autoanalyzer (Boehringer Mannheim, Indianapolis, USA). To examine the effect of the delay between blood withdrawal and electrolyte measurement and the loss of CO2 by exposure to the room air after bottle opening, the subjects were divided into group A (124 subjects) and group B (414 subject) whose electrolyte results were reported within and after 20 minutes, respectively, of blood withdrawal and the two groups were compared with each other for the electrolytes and anion gap. Anion gap was calculated by the formula, [Na+- (Cl-+TCO2)]. RESULTS: Compared with group A, TCO2 of group B decreased by 0.9 mmol/L (P<0.001) and the anion gap increased by 0.6 mmol/L (P=0.009). The reference interval of anion gap was established based on the group A at the value of 5.4-13.4 mmol/L. CONCLUSIONS: The reference interval of anion gap determined in this study was lower than the value commonly used and should be established with a minimum delay in electrolyte measurement and sample exposure to the air.

Genomic analysis of Vibrio vulnificus by Infrequent Restriction Site-Polymerase Chain Reaction / 대한임상미생물학회지

BACKGROUND: Vibrio vulnificus sepsis is one of the notifiable disease(Class 3) in Korea. It is usually acquired through the consumption of raw or undercooked seafood in summer. We studied the clinical findings of V. vulnificus septicemia and the genomic patterns of V. vulnificus isolates. METHODS: Seven patients with V. vulnificus septicemia were admitted to Hanyang University hospital from 1998 to 2002. We analysed the clinical findings and the genomic patterns by infrequent restriction site-polymerase chain reaction(IRS-PCR). RESULTS: All patients were over forty years old, and five were male. The patients had underlying diseases;five with liver cirrhosis, two with DM, and four patients with heavy alcoholism. Five of seven patients had history of ingesting raw fish and four had tissue necrosis with bullae or vesicles in their extremities. Four patients who died showed disseminated intravascular coagulation symptoms. We applied IRS-PCR to 6 isolates from blood and 2 isolates from wound. The six isolates from blood showed various genomic patterns that were all different from one another, while the two isolates from wound showed IRS-PCR patterns that were identical to the blood isolates of the same patients. CONCLUSIONS: The genomic patterns of IRS-PCR are quite different in 6 cases of V. vulnificus isolates in Korea.

Expression of insulin-like growth factorIGF-II and IGF binding proteinIGFBP-1 in preeclamptic placentas / 대한산부인과학회잡지

No abstract available.

Expression of Hepatocyte Growth Factor and its receptor in Placentas of Mild and Severe Preeclampsia / 대한산부인과학회잡지

No abstract available.