In order to explore the factors that affect CD62p expression in platelet during the whole course of cryopreservated platelets preparation, CD62p expression of platelet was evaluated by flow cytometry assay. The whole course of cryopreservated platelets preparation in order included whole blood collection, centrifugation for fresh platelet-rich plasma preparation, addition of dimethyl sulfoxide, and freeze in -80 degrees C refrigerator and thaw in 38 degrees C water bath. Result showed that the CD62p expressed slowly from whole blood collection to addition of dimethyl sulfoxide, but expressed abruptly after freeze and thaw and it occupied 82 per cent of the whole expression. It was concluded that the whole blood collection, centrifugation for fresh platelet enriched plasma preparation and addition of dimethyl sulfoxide were optimized in the whole course, but the damage to platelets in the whole course of cryopreservation could not be avoided. It suggests that improved techniques are needed to reduce the damage to cryopreservated platelets.
Blood Platelets ; metabolism ; Blood Preservation ; methods ; standards ; Cryopreservation ; methods ; standards ; Flow Cytometry ; Humans ; P-Selectin ; biosynthesis

BACKGROUND: This study aimed to analyze the influence of the interruption of agitation and removal of leukocytes on platelet concentrates (PCs), and determine the maximum amount of time the agitation could be interrupted without impairing PCs' effectiveness during the storage period. METHODS: Four ABO-identical random donor platelets agitated for 24 hr were pooled, and divided into 4 units, and 2 units of them were leukoreduced. Then 52 pooled units were categorized into 4 groups, non-leukoreduced continuous agitation (Non-LRCA), non-leukoreduced interrupted agitation (Non-LRIA), leukoreduced continuous agitation (LRCA), and leukoreduced interrupted agitation (LRIA), and preserved for 6 days (total 7 days). Mean platelet volume (MPV), pH, HCO3-, pO2, pCO2, CD62P, CD61, glucose, lactate, ammonia and free fatty acid were measured during the period. RESULTS: Starting from the Day 4, the pH and HCO3- of Non-LRIA group begun to decrease while the amount of lactate production, glucose consumption, and MPV increased compared to the Non- LRCA group (P<0.01). An increase in pO2 level was observed in the interrupted agitation groups as the storage period prolonged (P<0.01). The pH levels of all the units in the agitation groups remained higher than 6.4 up to Day 7, while those of the non-leukoreduction group did so only up to Day 2, but those of leukoreduction in the interrupted agitation groups did so up to Day 4. CONCLUSIONS: The interruption of agitation reduced the platelet's capacity to utilize oxygen, increasing lactate amount and reducing pH level. However, the in vitro parameters of the Non-LRIA and Non-LRCA groups on Day 2 were similar to each other and the pH level remained at 6.4 or higher, making one day of agitation interruption possible after 24 hr of agitation. With leukocytes removed, the effective agitation interruption period may become longer.
*Blood Component Removal ; Blood Platelets/*cytology ; Blood Preservation/*standards ; Cell Separation ; Glucose/analysis ; Humans ; Hydrogen-Ion Concentration ; Lactic Acid/blood ; Oximetry ; P-Selectin/blood ; Time Factors ; Vibration

The study was purposed to investigate whether processing and storage conditions might influence the stability of the HCV RNA in whole blood or in plasma. The samples obtained from seven patients known to be positive for HCV RNA were kept in different storage conditions with different anticoagulants, and at the end of processing the plasma samples were frozen at -80 degrees C until fluorescent quantitative PCR testing. The results showed that there was no significant loss of HCV RNA titers in whole blood anticoagulated with CPDA or ACD or EDTA or none (P > 0.05), while differences in comparison of the EDTA-anticoagulant storage condition with three other anticoagulants storage conditions at 4 degrees C after 48 hours were significant (P < 0.05). The HCV RNA level decreased to 53.8%, 72.5% and 29.8% after 48 hours of storage of whole blood anticoagulated with ACD at 4 degrees C, 25 degrees C and 37 degrees C respectively. The HCV RNA level of plasma samples stored at 4 degrees C and at 25 degrees C (room temperature) after 7 days decreased to 70.9% and 25.1% respectively. After four freeze-thaw cycles the HCV RNA level decreased 38.9% in plasma samples. It is concluded that the HCV RNA is stable relatively. The HCV RNA is resistant to degradation under routine laboratory handling and storage conditions or blood collection, transport and processing conditions. The influence of different anticoagulants on the stability of HCV RNA is different. Blood samples would better be stored at 4 degrees C after collection and plasma separated within 48 hours. And it is important for the stability of HCV RNA undergoing asepsis blood collection process. HCV RNA remains stable at 4 degrees C for at least 7 days or at room temperature for 3 days, allowing greater flexibility in samples collection and transport in transfusion practice nowadays. HCV RNA in plasma samples subject to up to three short-term freeze-thaw cycles is still stable.
Blood Donors ; Blood Preservation ; methods ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; RNA, Viral ; blood ; drug effects ; Specimen Handling ; standards ; Temperature ; Time Factors

This study was purposed to investigate the effectiveness of bacterial screening with 24 hours holding in preventing and controlling bacterial contamination of platelets. Bacterial screening of apheresis platelets preserved for 24 hours was performed by using BacT/ALERT automatic bacterial culture system. The samples from 5 bags of platelet were taken in aseptic condition and were merged into 1 bag. The final sample was inoculated into aerobic and anaerobic bottle respectively for testing, meanwhile the screened platelet samples were held for 24 hours. If the platelets were cultured for 24 hours and identification of bacterial strains showed negative, the platelets could be released, and the original platelet samples should be rescreened if initiate positive was found. The results showed that in screening 8017 samples of apheresis platelets the initiate positive results were 16 (0.2%) and confirmed positive were 4 (0.05%). Out of 4 confirmed positive strains, three were Staphylococcus aureus and another was Staphylococcus auricularis. It is concluded that it is necessary for blood center to apply the method of bacterial screening of platelet with 24 hours holding as conventional screening method, which is an effective and feasible way to prevent and control bacterial contamination of platelets.
Bacteria ; isolation & purification ; Bacterial Infections ; prevention & control ; Bacteriological Techniques ; instrumentation ; Blood Platelets ; microbiology ; Blood Preservation ; methods ; standards ; Humans ; Platelet Transfusion ; adverse effects ; Plateletpheresis ; instrumentation

BACKGROUND: The red blood cell (RBC) deformability test is a useful method for measuring the ability of RBCs to adapt their shape to the flow conditions. Using this test, several investigators have shown the relationship between RBC deformability and numerous clinical conditions. For the quality control (QC) of RBC deformability test, we evaluated whether frozen-thawed-deglycerolized RBCs can be used as QC materials. METHODS: Packed RBCs were frozen with 40% (wt/vol) glycerol and stored at -80degrees C for 3 months. For 10 different frozen RBC panels, RBCs were thawed, deglycerolized and stored at 4degrees C for 4 weeks. Using microfluidic ektacytometer, we measured RBC deformability of the thawed RBCs. The stability of thawed RBCs was tested once a day for 28 days of storage time and was analyzed by simple regression analysis. The precision of the test using thawed RBCs was analyzed for 7 days of storage time by calculation of CV values of intra-assay (10 measurements/assay) and between-day measurements. RESULTS: Frozen-thawed-deglycerolized RBCs were stable for 1 week. Within-run and between-day precisions of the RBC deformability test during 7 days of storage of thawed RBCs were 1.4-2.9%, and 1.9-2.8%, respectively. CONCLUSIONS: Frozen-thawed-deglycerolized RBCs used in RBC deformability test showed satisfactory within-run and between-run precisions and stability for one week after thawing, and may be used as QC materials for this test.
Blood Preservation ; Cryopreservation ; Cryoprotective Agents/chemistry/isolation & purification ; *Erythrocyte Deformability ; Erythrocytes/immunology/physiology ; Glycerol/chemistry/isolation & purification ; Hematologic Tests/standards ; Humans ; Quality Control

INTRODUCTIONIntraoperative cell salvage (ICS) is an important aspect of patient blood management programmes. An ICS service was introduced at KK Women's and Children's Hospital, Singapore, from 2 May 2011 to 30 April 2013 to aid in the management of massive obstetric haemorrhage.

METHODSWith support from the Ministry of Health's Healthcare Quality Improvement and Innovation Fund, a workgroup comprising obstetricians, anaesthetists and nursing staff was formed to develop training requirements, clinical guidelines and protocols for implementing ICS using the Haemonetics Cell Saver 5. Pregnant women with an anticipated blood loss of > 1,000 mL during Caesarean delivery, a baseline haemoglobin level of < 10 g/dL, rare blood types and who had refused donor blood were recruited to the service after obtaining informed consent.

RESULTSA total of 11 women were recruited to the ICS service; the primary indications were placenta praevia and placenta accreta. Median blood loss in these 11 patients was 1,500 (range 400-3,000) mL. In four patients, adequate autologous blood was collected to initiate processing and salvaged, processed blood was successfully reinfused (mean 381.3 [range 223.0-700.0] mL). Median blood loss among these four patients was 2,000 (range 2,000-3,000) mL. No adverse event occurred following autologous transfusion. Mean immediate postoperative haemoglobin level was 8.0 (range 7.1-9.4) g/dL.

CONCLUSIONThe implementation of an obstetric ICS service in our institution was successful. Future studies should seek to address the cost-effectiveness of ICS in reducing allogeneic blood utilisation.

Blood Preservation ; Blood Transfusion, Autologous ; methods ; standards ; Cost-Benefit Analysis ; Female ; Hemoglobins ; analysis ; Hemorrhage ; therapy ; Humans ; Obstetrics ; methods ; standards ; Operative Blood Salvage ; methods ; standards ; Placenta Accreta ; therapy ; Placenta Previa ; therapy ; Practice Guidelines as Topic ; Pregnancy ; Program Development ; Program Evaluation ; Singapore ; Tertiary Care Centers