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  • Original Article2022-11-01 Transfusion and Cell Therapy

    Natural Killer Cell Expansion and Cytotoxicity Differ Depending on the Culture Medium Used

    Seung Kwon Koh , B.S., Jeehun Park , Ph.D., Seong-Eun Kim , B.S., Yuree Lim , M.S., Minh-Trang Thi Phan , Ph.D., Jinho Kim , B.S., Ilwoong Hwang , M.D., Yong-Oon Ahn , Ph.D., Sue Shin , M.D., Junsang Doh , Ph.D., and Duck Cho , M.D.

    Ann Lab Med 2022; 42(6): 638-649

    Abstract : Background: Adoptive cell therapy using umbilical cord blood (UCB)-derived allogeneic natural killer (NK) cells has shown encouraging results. However, because of the insufficient availability of NK cells and limited UCB volume, more effective culture methods are required. NK cell expansion and functionality are largely affected by the culture medium. While human serum is a major affecting component in culture media, the way it regulates NK cell functionality remains elusive. We elucidated the effects of different culture media and human serum supplementation on UCB NK cell expansion and functionality. Methods: UCB NK cells were cultured under stimulation with K562-OX40L-mbIL-18/21 feeder cells and IL-2 and IL-15 in serum-containing and serum-free culture media. The effects of the culture media and human serum supplementation on NK cell expansion and cytotoxicity were evaluated by analyzing the expansion rate, activating and inhibitory receptor levels, and the cytotoxicity of the UCB NK cells. Results: The optimal medium for NK cell expansion was Dulbecco’s modified Eagle’s medium/Ham’s F12 with supplements and that for cytotoxicity was AIM V supplemented with Immune Cell Serum Replacement. Shifting media is an advantageous strategy for obtaining several highly functional UCB NK cells. Live cell imaging and killing time measurement revealed that human serum enhanced NK cell proliferation but delayed target recognition, resulting in reduced cytotoxicity. Conclusions: Culture medium supplementation with human serum strongly affects UCB NK cell expansion and functionality. Thus, culture media should be carefully selected to ensure both NK cell quantity and quality for adoptive cell therapy.

  • Original Article2022-07-01 Transfusion and Cell Therapy

    Erythroid Differentiation of Induced Pluripotent Stem Cells Co-cultured with OP9 Cells for Diagnostic Purposes

    Juhye Roh , M.D., Ph.D., Sinyoung Kim , M.D., Ph.D., June-Won Cheong , M.D., Ph.D., Su-Hee Jeon , M.S., Hyun-Kyung Kim , M.S., Moon Jung Kim , M.D., Ph.D., and Hyun Ok Kim , M.D., Ph.D.

    Ann Lab Med 2022; 42(4): 457-466

    Abstract : Background: Reagent red blood cells (RBCs) are prepared from donated whole blood, resulting in various combinations of blood group antigens. This inconsistency can be resolved by producing RBCs with uniform antigen expression. Induced pluripotent stem cells (iPSCs) generated directly from mature cells constitute an unlimited source for RBC production. We aimed to produce erythroid cells from iPSCs for diagnostic purposes. We hypothesized that cultured erythroid cells express surface antigens that can be recognized by blood group antibodies. Methods: iPSCs were co-cultured with OP9 stromal cells to stimulate differentiation into the erythroid lineage. Cell differentiation was examined using microscopy and flow cytometry. Hemoglobin electrophoresis and oxygen-binding capacity testing were performed to verify that the cultured erythroid cells functioned normally. The agglutination reactions of the cultured erythroid cells to antibodies were investigated to confirm that the cells expressed blood group antigens. Results: The generated iPSCs showed stemness characteristics and could differentiate into the erythroid lineage. As differentiation progressed, the proportion of nucleated RBCs increased. Hemoglobin electrophoresis revealed a sharp peak in the hemoglobin F region. The oxygen-binding capacity test results were similar between normal RBCs and cultured nucleated RBCs. ABO and Rh-Hr blood grouping confirmed similar antigen expression between the donor RBCs and cultured nucleated RBCs. Conclusions: We generated blood group antigen-expressing nucleated RBCs from iPSCs co-cultured with OP9 cells that can be used for diagnostic purposes. iPSCs from rare blood group donors could serve as an unlimited source for reagent production.

  • Original Article2022-11-01 Transfusion and Cell Therapy

    Investigation of Discrepant ABO Blood Grouping Results from an Autoanalyzer

    Young Ae Lim , M.D., Ph.D., Seo-Jin Park , M.D., Ph.D., and Hyun Soo Cho , M.S.

    Ann Lab Med 2022; 42(6): 650-658

    Abstract : Background: A paucity of studies evaluating failed cases of ABO grouping using autoanalyzers exists. We investigated autoanalyzer rejected cases, including serologically suspicious ABO subgroups and discrepant ABO blood grouping results from Erytra Eflexis (Grifols, Spain), to demonstrate efficient use of autoanalyzers for ABO grouping. Methods: Samples requested for ABO grouping throughout 2020 were tested using two Eflexis instruments and standard ABO RhD and reverse grouping cards. Neonatal cards were not used. When necessary, a conventional tube technique (TUBE) was used to resolve rejected/discrepant Eflexis ABO grouping results. Results: The overall sample rejection rate (RR) was 3.2% (628/19,466), 1.3% of which were due to various error flags and 1.9% for discrepant results. Cases from neonates ≤1 year old accounted for 35.3% of the rejected cases based on Eflexis results. The ABO groups with the highest and lowest RR (excluding neonates) were A and O, respectively. The 628 samples resulted in 682 rejections, which were frequently associated with reverse grouping, including 28.4% against A1 and 54.5% for B red cells. Among 14 serologically weakened A and/or B blood groups, six A2BW and two ABw, which had been missed by Eflexis, were detected using TUBE and our follow-up laboratory criteria. Conclusions: The ABO group and a proportion of neonatal samples influenced the RR due to weak reverse grouping reactivity, especially toward B red cells. Confirmatory ABO grouping by TUBE in a new patient and/or extra rejection criteria for forward grouping are needed to detect cis-AB, which is relatively common in Korea.

  • Brief Communication2023-01-01 Transfusion and Cell Therapy

    Improvement of Anti-CD36 Antibody Detection via Monoclonal Antibody Immobilization of Platelet Antigens Assay by Using Selected Monoclonal Antibodies

    Xiuzhang Xu , Ph.D., Dawei Chen , Ph.D., Xin Ye , M.D., Wenjie Xia , Ph.D., Yuan Shao , Ph.D., Jing Deng , M.T., Yangkai Chen , M.T., Haoqiang Ding , M.T., Jing Liu , M.S., Yaori Xu , M.T., Sentot Santoso , Ph.D., and Yongshui Fu , M.D.

    Ann Lab Med 2023; 43(1): 86-91

    Abstract : Antibodies against human CD36 are responsible for several immune-mediated disorders. The detection of anti-CD36 antibodies using the standard monoclonal antibody (mAb) immobilization of platelet antigens (MAIPA) assay is hampered by a high frequency of false-negative results, most likely due to competitive inhibition of the mAb used as the capture antibody. We generated a panel of mouse mAbs against CD36 and seven hybridomas (GZ-3, GZ-13, GZ-70, GZ-143, GZ-413, GZ-507, and GZ-608), which were selected for MAIPA assays, as they reacted with mouse and human CD36. Fourteen anti-CD36 sera were assayed; all of which showed a positive reaction in a PakPlus (Immucor GTI Diagnostics, Inc., Waukesha, WI, USA) ELISA-based screening (optical density: 0.257–2.292). When the reference anti-CD36 mAb FA6-152 was used in the MAIPA assay, only 6/14 (42.9%) sera displayed a positive reaction. In contrast, anti-CD36 antibodies were detected in 13/14 (92.9%) sera when GZ-70 and GZ-608 mAbs were used. This significant improvement resulted in the identification of anti-CD36 antibodies by an antigen capture assay. Since patient’s platelets possibly carrying rare native antigens are used, this method will facilitate the identification of new platelet antibodies against CD36 that are involved in immune-mediated thrombocytopenia and other diseases, such as transfusion-related acute lung injury.

  • Original Article2022-11-01 Transfusion and Cell Therapy

    Predictors of Red Blood Cell Transfusion in Elderly COVID-19 Patients in Korea

    Hye Ryun Lee , M.D., Ph.D.

    Ann Lab Med 2022; 42(6): 659-667

    Abstract : Background: Patients who experience clinical deterioration from coronavirus disease (COVID-19) require blood transfusion support. We analyzed blood component usage in COVID-19 patients and identified the predictors of red blood cell (RBC) transfusion in elderly (≥65 years) patients. Methods: Blood component usage in 882 COVID-19 patients hospitalized between January 24, 2020 and April 30, 2021 was analyzed. Elderly patients were categorized into transfused and non-transfused groups according to their RBC transfusion history; their demographic and clinical characteristics, disease severity, and outcomes were compared. Associations were determined using multiple logistic regression. Results: The overall transfusion rate was 8.3% (73/882), and the transfusion rate was 2.7% (14/524) in patients aged

  • Original Article2023-03-01 Transfusion and Cell Therapy

    Common Data Model-based Analysis of Selective Leukoreduction Protocol Compliance at Three Hospitals

    Sooin Choi , M.D., M.S., Soo Jeong Choi , M.D., Ph.D., Jeong Won Shin , M.D., Ph.D., and Young Ahn Yoon , M.D., Ph.D.

    Ann Lab Med 2023; 43(2): 187-195

    Abstract : Background: The selective leukoreduction protocol (SLP) is limited in that patients who require it can be overlooked. We estimated SLP compliance (SLPC) using the Observational Medical Outcomes Partnership common data model (CDM). Methods: Patients were classified into eight groups: pre- and post-hematology disease (A and B), pre- and post-solid organ transplantation (C and D), solid cancer (E), immunodeficiency (F), anticancer therapy (G), and cardiovascular surgery (H). We examined the red blood cell (RBC) transfusion history from three hospital datasets comprising approximately three million patients over 20 years using CDM-based analysis. SLPC was calculated as the percentage of patients who received only leukoreduced RBCs in total patients transfused RBCs. Results: In total, 166,641 patients from three hospitals were enrolled in this study. From 2001 to 2021, SLPC in all groups, except H, tended to increase, although there were differences among the hospitals. Based on the most recent values (2017–2021), the SLPC in groups A, B, D, and G was maintained at ≥75% until 1,095 days before or after diagnosis or treatment. Groups E, F, and H had < 50% SLPC one day after diagnosis and treatment. Conclusions: CDM analysis supports the review of large datasets for SLPC evaluation. Although SLPC tended to improve in most patient groups, additional education and monitoring are needed for groups that continue to show low SLPC.

  • Letter to the Editor2022-11-01 Transfusion and Cell Therapy

    Clinical Significance of Hemagglutination Grades Determined Using the IH-1000 Automated Blood Typing Instrument: Real-world Data

    Bosung Park , M.D., Jin Seok Kim , M.T., Hee-Jeong Youk , M.D., Yousun Chung , M.D., Hyungsuk Kim , M.D., Sang-Hyun Hwang , M.D., Heung-Bum Oh , M.D., and Dae-Hyun Ko , M.D.

    Ann Lab Med 2022; 42(6): 700-702
  • Original Article2023-09-01 Transfusion and Cell Therapy

    Performance Validation of Three Scoring Systems for the Prediction of Thrombotic Microangiopathy Due to Severe ADAMTS13 Deficiency and the Response to Therapeutic Plasma Exchange: First Study in Korea

    Sang Hyuk Park , M.D., Ph.D., Hyun-Ki Kim , M.D., Joseph Jeong , M.D., Ph.D., Seon-Ho Lee , M.D., Ph.D., Yoo Jin Lee , M.D., Ph.D., Yoo Jin Kim , M.D., Ph.D., Jae-Cheol Jo , M.D., Ph.D., and Ji-Hun Lim , M.D., Ph.D.

    Ann Lab Med 2023; 43(5): 485-492

    Abstract : Background: The BENTLEY score (B-S), French thrombotic microangiopathy (TMA) Reference Center score (FTMA-S), and PLASMIC score (PLASMIC-S) have been developed for TMA diagnostic prediction. We retrospectively validated their predictive performances in patients with severe (

  • Letter to the Editor2022-11-01 Transfusion and Cell Therapy

    Proposal of the Need for New Korean Guidelines on the Use of Therapeutic Apheresis in Clinical Practice

    Yousun Chung , M.D., Hyungsuk Kim , M.D., and Dae-Hyun Ko , M.D.

    Ann Lab Med 2022; 42(6): 703-707
  • Original Article2023-09-01 Transfusion and Cell Therapy

    Reducing Microbial Contamination in Hematopoietic Stem Cell Products and Quality Improvement Strategy: Retrospective Analysis of 1996-2021 Data

    You Keun Ko , M.D., Jong Kwon Lee , M.D., Hye Kyung Park , M.T., Ae Kyung Han , M.T., Sun Kyoung Mun , M.T., Hye Jeong Park , M.T., Hae Kyoung Choung , R.N., Se Mi Kim , R.N., Kwang Mo Choi , M.T., Nam Yong Lee , M.D., Ph.D., Duck Cho , M.D., Ph.D., Dae Won Kim , M.D., Ph.D., and Eun-Suk Kang , M.D., Ph.D.

    Ann Lab Med 2023; 43(5): 477-484

    Abstract : Background: Sterility and safety assurance of hematopoietic stem cell (HSC) products is critical in transplantation. Microbial contamination can lead to product disposal and increases the risk of unsuccessful clinical outcomes. Therefore, it is important to implement and maintain good practice guidelines and regulations for the HSC collection and processing unit in each hospital. We aimed to share our experiences and suggest strategies to improve the quality assurance of HSC processing. Methods: We retrospectively analyzed microbial culture results of 11,743 HSC products processed over a 25-year period (January 1996 to May 2021). Because of reorganization of the HSC management system in 2008, the 25-year period was divided into periods 1 (January 1996 to December 2007) and 2 (January 2008 to May 2021). We reviewed all culture results of the HSC products and stored aliquot samples and collected culture results for peripheral blood and catheter samples. Results: Of the 11,743 products in total, 35 (0.3%) were contaminated by microorganisms, including 19 (0.5%) of 3,861 products during period 1 and 16 (0.2%) of 7,882 products during period 2. Penicillium was the most commonly identified microorganism (15.8%) during period 1 and coagulase-negative Staphylococcus was the most commonly identified (31.3%) during period 2. HSC product contamination occurred most often during HSC collection and processing. Conclusions: The contamination rate decreased significantly during period 2, when the HSC management system was reorganized. Our results imply that handling HSC products by trained personnel and adopting established protocols, including quality assurance programs, aid in decreasing the contamination risk.

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Annals of Laboratory Medicine
Journal Information May, 2024
Vol.44 No.3
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