Evaluation of kodecytes using function-spacer-lipid constructs as a survey material for external proficiency testing for ABO subgrouping.

BACKGROUND: It is not easy to find natural red blood cells (RBCs) with weak A (Aw ) or weak B phenotype (Bw ) for use as quality controls in ABO subgroup testing (subgrouping). The aim of this study was to prepare RBC kodecytes with synthetic blood group A and/or B function-spacer-lipid (FSL) constructs and to evaluate the possibility of using such kodecytes as a survey material for an external proficiency test (PT) to improve the quality of subgroup analysis. METHODS: Three types of survey samples, including O phenotype RBCs and A kodecytes with Aw (0.02 mg/mL FSL-A solution) and B kodecytes with Bw (0.15 mg/mL FSL-B solution) were sent to 53 laboratories for an educational trial of PT for subgrouping. Cell typing was done using the manual tube technique. RESULTS: Forty-three laboratories responded, and the re-activities of the survey samples varied from 0 to 4+ against anti-A and anti-B monoclonal reagents(MoAbs). Twenty-nine laboratories (67%) correctly grouped the Bw kodecytes as Bw . Fifteen (35%), 21 (48%), and 6 (13%) laboratories grouped the Aw kodecytes as Aw , A2 , and O phenotypes, respectively. The anti-A MoAb clone affects the results of cell typing for Aw kodecytes. The stability of kodecytes was similar to that of natural O RBCs during storage. CONCLUSION: Our kodecytes were useful as a survey material, and the survey results showed the necessity of materials for PT for subgrouping to improve the quality of laboratory analysis regardless of the different reactions according to the MoAb.

Standardization of ABO antibody titer measurement at laboratories in Korea.

BACKGROUND: Measurement of the ABO antibody (Ab) titer is important in ABO-incompatible transplantation. However, to the best of our knowledge, no standard protocol or external survey program to measure the ABO Ab titer has been established in Korea. We investigated the current status of ABO Ab titer measurements at various laboratories in Korea and the impact of the protocol provided to reduce interlaboratory variations in the methods and results of ABO Ab titers. METHODS: The Korean external quality assessment of blood bank laboratories sent external survey samples with a questionnaire to 68 laboratories across Korea for the measurement of ABO Ab titers in May 2012. After 6 months, a second set of survey samples were sent with a standard protocol to 53 of the previously surveyed laboratories. The protocol recommended incubation at room temperature only and use of the indirect antihuman globulin method for the tube test as well as and the column agglutination test (CAT). RESULTS: Several interlaboratory variations were observed in the results, technical procedures, and methods selected for measurement. We found that 80.4% laboratories hoped to change their protocol to the provisional one. Additionally, CAT showed significantly lower variation among laboratories (P=0.006) than the tube test. CONCLUSIONS: Our study provides baseline data regarding the current status of ABO Ab titer measurement in Korea. The standard protocol and external survey were helpful to standardize the technical procedures and select methods for ABO Ab titer measurement.

Comparison of ABO antibody titers on the basis of the antibody detection method used.

BACKGROUND: Detection methods for ABO antibody (Ab) titers vary across laboratories, and the results are different depending on the method used. We aimed to compare titer values using different detection methods for the measurement of ABO Ab titers. METHODS: For ABO Ab detection, pooled group A or B red blood cells (RBCs) were reacted with each of 20 sera from blood groups A, B, or O without dithiothreitol treatment. The room-temperature (RT) incubation technique and the indirect antiglobulin test (IAT) were used in the tube test and gel card test. Flow cytometry (FCM) was performed by using anti-IgM and anti-IgG Abs. RESULTS: Regardless of the blood groups tested, the FCM assay with anti-IgM showed the highest titer compared to the tube test and gel card test with RT incubation in both. The tube test with IAT showed a higher titer than the gel card test with IAT (Gel-IAT) or FCM with anti-IgG in blood group A and B, while Gel-IAT showed the highest titer relative to the other tests, only for the anti-A Ab in blood group O. CONCLUSIONS: There were significant differences in the titers depending on the detection method used, and each method showed a different detection capacity for each ABO Ab depending on the ABO blood group tested. Therefore, caution should be exercised in interpreting ABO Ab titer results, taking into consideration the detection method used and the blood group.

Performance evaluation of Samsung LABGEO(HC10) Hematology Analyzer.

CONTEXT: The Samsung LABGEO(HC10) Hematology Analyzer (LABGEO(HC10)) is a recently developed automated hematology analyzer that uses impedance technologies. The analyzer provides 18 parameters including 3-part differential at a maximum rate of 80 samples per hour. OBJECTIVE: To evaluate the performance of the LABGEO(HC10). DESIGN: We evaluated precision, linearity, carryover, and relationship for complete blood cell count parameters between the LABGEO(HC10) and the LH780 (Beckman Coulter Inc) in a university hospital in Korea according to the Clinical and Laboratory Standards Institute guidelines. Sample stability and differences due to the anticoagulant used (K2EDTA versus K3EDTA) were also evaluated. RESULTS: The LABGEO(HC10) showed linearity over a wide range and minimal carryover (<1%) for white blood cell, hemoglobin, red blood cell, and platelet parameters. Correlation between the LABGEO(HC10) and the LH780 was good for all complete blood cell count parameters (R > 0.92) except for mean corpuscular hemoglobin concentration. The bias estimated was acceptable for all parameters investigated except for monocyte count. Most parameters were stable until 24 hours both at room temperature and at 4°C. The difference by anticoagulant type was statistically insignificant for all parameters except for a few red cell parameters. CONCLUSIONS: The accurate results achievable and simplicity of operation make the unit recommendable for small to medium-sized laboratories.

Effects of platelet lysate preparations on the proliferation of HaCaT cells.

BACKGROUND: Standard protocols are lacking for the preparation of platelet lysates (PL) as an alternative to using fetal bovine serum as a cell culture supplement. This study aimed to establish optimum conditions for preparing PL for use in cell cultures. METHODS: Cell density in three pooled platelet concentrates (PC) were adjusted to 1×10(12)/L and 2×10(12)/L. PL was prepared from PC by 1 to 3 freeze-thaw (FT) cycles. HaCaT cells were cultured in media supplemented with 5% or 10% PL. Cell numbers were estimated using a Cell Counting Kit-8 (CCK-8; Dojindo Laboratories, Japan). Growth factors were quantified by using the Luminex 200 system (Luminex Corporation, USA). RESULTS: Cell proliferation rates in the presence of PLs were similar when prepared from PCs of both cell densities. The rates were higher in media containing 5% PL than 10% PL when prepared by two FT cycles. Concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor-AB/BB (PDGF-AB/BB), PDGF-AA, and epidermal growth factor (EGF) were significantly higher in PL prepared from PC with a cell density of 2×10(12)/L than 1×10(12)/L PC. However, only VEGF and PDGF-AA concentrations in PLs were correlated with HaCaT cell counts. CONCLUSIONS: The 5% PL from PC with a cell density of 1×10(12)/L prepared by two FT cycles treatment was the most effective condition that supported steady HaCaT cell proliferation. Our finding may be useful for preparing PL-supplemented cell culture media.