Comparison of automated and manual methods for washing red blood cells.

BACKGROUND: We sought to compare the quality of washed red blood cells (RBCs) produced using the ACP215 device or manual methods with different combinations of wash and storage solutions. Our aim was to establish manual methods of washing that would permit a shelf life of more than 24 hours. STUDY DESIGN AND METHODS: Fourteen-day-old RBCs were pooled, split, and washed in one of five ways: 1) using the ACP215 and stored in SAGM, 2) manually washed and stored in saline, 3) manually washed in saline and stored in SAGM, 4) manually washed in saline-glucose and stored in SAGM, and 5) manually washed and stored in SAGM. Additional units were pooled and split, washed manually or using the ACP215, and irradiated on Day 14. Units were sampled to 14 days after washing and storage at 4 ± 2°C. RESULTS: All washed RBCs met specification for volume (200-320 mL) and hemoglobin (Hb) content (>40 g/unit). Removal of plasma proteins was better using manual methods: residual immunoglobulin A in saline-glucose-washed cells 0.033 (0.007-0.058) mg/dL manual versus 0.064 (0.026-0.104) mg/dL ACP215 (median, range). Hb loss was lower in manually washed units (mean, ≤ 2.0g/unit) than in ACP215-washed units (mean, 6.1 g/unit). Disregarding saline-washed and stored cells, hemolysis in all nonirradiated units was less than 0.8% 14 days after washing. As expected, the use of SAGM to store manually washed units improved adenosine triphosphate, glucose, lactate, and pH versus storage in saline. CONCLUSION: The data suggest that the shelf life of manually washed RBCs could be extended to 14 days if stored in SAGM instead of saline.

Platelet storage in more than 90% additive solution containing glucose and bicarbonate has the potential to increase shelf life.

BACKGROUND: Platelet concentrates (PCs) suspended in more than 90% additive solution (AS) are recommended for patients with reactions to platelets stored in plasma. Next-generation AS containing glucose and bicarbonate might enable storage of these PCs for longer than those in current-generation AS, which was therefore evaluated in this study. STUDY DESIGN AND METHODS: Five buffy coat or apheresis-derived PCs were pooled and split into identical units. All units except the control were centrifuged, the plasma removed and replaced with AS (SSP+, PAS-5, or PAS-G), or resuspended in the same plasma and sampled 96 hours after resuspension for analysis. RESULTS: Plasma carryover was less than 10%, total protein less than 1 g/unit, and immunoglobulin A levels lower than 0.1 mg/mL for all PCs in AS. The pH of all the platelets during storage was higher than 6.4. PAS containing glucose maintained superior in vitro platelet function during storage compared with those resuspended in SSP+. Compared with storage in SSP+, storage in PAS-5 or PAS-G resulted in better preservation of platelet adenosine triphosphate and hypotonic shock response, lower annexin V binding, and improved mitochondrial membrane potential. CONCLUSION: Platelets resuspended in PAS-5 and PAS-G maintained in vitro function and metabolism during storage compared with SSP+ platelets. Elevated platelet metabolic activity was noticed in PAS-G, and higher platelet activation was detected with PAS-5. Platelets resuspended in PAS containing glucose has the potential to increase the shelf life of PC in more than 90% AS.

Fast Track to Accreditation: An Implementation Review of College of American Pathologists and International Organization for Standardization 15189 Accreditation.

CONTEXT: - This review examines challenges and opportunities in preparing laboratories in a startup phase for accreditation by both the College of American Pathologists (CAP) and International Organization for Standardization (ISO) 15189 in an international setting as it relates to our experience at Cleveland Clinic Abu Dhabi Laboratory. It also discusses some of the strategies used in executing those projects and the added advantages in pursuing both types of accreditations. OBJECTIVES: - To share our experience with CAP and ISO 15189 accreditations in a startup international operation in relation to the challenges encountered and implementation strategy success factors. DATA SOURCES: - MEDLINE (PubMed) database was used to review this topic as well as peer-reviewed articles and World Health Organization publications on the topic. CONCLUSIONS: - Accreditation is a perfect means toward building quality medical laboratories in a diverse workforce environment and improving patient safety. Further, it establishes a strong foundation on which any new operation can build a sustainable quality improvement culture. Accreditations by CAP and/or ISO are among the most reputable and well-established accreditation systems that clinical laboratories could aim for. As a result of both accreditations offering synergistic and complementing features, we recommend that any laboratory seeking excellence in quality and performance should consider exploring both. Key elements to success include having dedicated project management and change management support while preparing for accreditation. Laboratories seeking accreditation in early operational stages may face a number of challenges; however, significant opportunities will also be present to optimize various operational components from the beginning.

Washing red cells after leucodepletion does not decrease human leukocyte antigen sensitization risk in patients with chronic kidney disease.

BACKGROUND: Standard leucodepleted blood transfusions can induce the production of human leukocyte antigen (HLA)-specific antibodies, which are associated with longer transplant waiting times and poorer graft outcomes. We hypothesized that additional washing of leucodepleted red cells might reduce antigenic stimulus by removal of residual leukocytes and soluble HLA. METHODS: A retrospective review of HLA antibodies in children with chronic kidney disease stage 4-5 who had ≥ two HLA antibody screens between 2000 and 2009, pre- and post-transfusion, and were HLA antibody-negative at first testing. Patients were divided according to whether they received standard leucodepleted blood or "washed cells". To assess the efficacy of washing methods, total leukocytes were enumerated pre- and post- manual and automated washing of standard leucodepleted red cells that had been supplemented with whole blood to achieve measurable leukocyte levels pre-washing. RESULTS: A total of 106 children were included: 23 received no blood transfusions (group 1), six had washed cells only (group 2), 59 had standard transfusions only (group 3), and 18 had both standard and washed cells (group 4). Sensitization rates were 26, 17, 44, and 44 % in groups 1-4 (p = 0.32). Patients in groups 3 and 4 had more transfusions with red cells, platelets, and plasma products. There was no difference in HLA sensitization risk with washed or standard red cells on analysis of co-variance controlling for platelets and plasma transfusions. The red cell washing study showed no significant reduction in leukocytes using manual methods. Although there was a statistically significant reduction (33 %) from baseline pre-washing using the automated method, from 6.54 ± 0.84 × 10(6) to 4.36 ± 0.67 × 10(6) leukocytes per unit, the majority of leukocytes still remained. CONCLUSIONS: There was no evidence that using washed leucodepleted red cells reduced patient HLA sensitization rates. Washing leucodepleted red cells is unlikely to reduce the risk of HLA sensitization due to the limited effect on residual leukocytes.

Pathogen inactivation of platelets using ultraviolet C light: effect on in vitro function and recovery and survival of platelets.

BACKGROUND: We evaluated the effect of treating platelets (PLTs) using ultraviolet (UV)C light without the addition of any photosensitizing chemicals on PLT function in vitro and PLT recovery and survival in an autologous radiolabeled volunteer study. STUDY DESIGN AND METHODS: For in vitro studies, pooled or single buffy coat-derived PLT concentrates (PCs) were pooled and split to obtain identical PCs that were either treated with UVC or untreated (n = 6 each) and stored for 7 days. PLT recovery and survival were determined in a two-arm parallel autologous study in healthy volunteers performed according to BEST guidelines. UVC-treated or untreated PCs (n = 6 each) were stored for 5 days and were compared to fresh PLTs from the same donor. RESULTS: There were no significant differences on Day 7 of storage between paired UVC-treated and control PC units for pH, adenosine triphosphate, lactate dehydrogenase, CD62P, CD63, PLT microparticles, and JC-1 binding, but annexin V binding, lactate accumulation, and expression of CD41/61 were significantly higher in treated units (p < 0.05). Compared with control units, the recovery and survival of UVC-treated PC were reduced after 5 days of storage (p < 0.05) and when expressed as a percentage of fresh values, survival was reduced by 20% (p = 0.005) and recovery by 17% (p = 0.088). CONCLUSION: UVC-treated PLTs stored for 5 days showed marginal changes in PLT metabolism and activation in vitro and were associated with a degree of reduction in recovery and survival similar to other pathogen inactivation systems that are licensed and in use.

Neutrophil function is preserved in a pooled granulocyte component prepared from whole blood donations.

Whole blood-derived granulocytes (buffy coats) are issued as an alternative to apheresis donations, but are heavily contaminated with red cells and platelets and there is minimal in vitro data describing their functionality. We developed a purer pooled granulocyte component (PGC) from whole blood donations by pooling 10 ABO-matched buffy coats with 400 ml of platelet additive solution (SSP+) and re-centrifuging. The PGC was irradiated (25-50 Gy) and neutrophil viability, chemotaxis, phagocytosis and respiratory burst activity were determined by flow cytometry. Results from 13 PGC at 16-20 h following donation were compared with those obtained from 20 standard individual buffy coats and with fresh whole blood. The PGC contained similar numbers of neutrophils (approximately 0.9 x 10(10)) with a reduced volume and haemoglobin content when compared with 10 individual buffy coats. Neutrophils in the PGC maintained >90% viability, oxidative burst and phagocytic activity and their ability to migrate towards a chemoattractant 16-20 h following donation, which is similar to results obtained with either fresh whole blood or standard buffy coats. Therefore, neutrophil function in the PGC was preserved 16-20 h following donation, but this product had significantly lower red cell contamination compared with 10 buffy coats, which are currently transfused.

A platelet quality assessment scheme for comparing the performance of quality monitoring laboratories in the UK National Blood Service.

This exercise focused on performance of NBS quality monitoring establishments with respect to enumeration of low leucocyte and other quality indexes of platelet concentration. Paired identical leucodepleted platelet samples, spiked with WBC (20 cells/microl) in 'vacuette' or 'pouch' were assessed by participants (n = 20) on days 1, 2 and 5. For low WBC counting, all laboratories gave estimates within acceptable range (+/-25%) and good agreement between storage and assay methods was observed on days 1 and 2. Day 5 results showed greater variability. Under improved performance criteria (+/-15%), only one laboratory under-estimated at days 1 and 2. Similarly, other parameters demonstrated good agreement between storage methods on days 1 and 2. At day 5, mean results were often significantly different to previous days. Improved performance target (+/-15%) will allow identification of non-conformers.