New trends in the preparation and storage of platelets.
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Biomedical subjects
Publications and source records attributed to C F Högman.
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The mechanism for the transmission of Yersinia enterocolitica in blood components has been studied experimentally. One hypothesis is that, during a Yersinia infection in the blood donor, bacteria are phagocytosed by white cells (WBCs), but are not killed. After collection of blood from such a donor and component production, the bacteria are present in WBCs for some time, during which the unit appears sterile. Later, when the WBCs disintegrate, the bacteria are released and multiply in the unit. Aliquots of whole blood and buffy coat were inoculated with 100 colony-forming units (CFU) per mL of a Y. enterocolitica strain of type O:3 and left at room temperature for 5 hours. Some aliquots were then WBC-reduced by filtration, while others retained their WBC contents. All aliquots were kept at 4 degrees C for 6 weeks. Meat extract broth culture medium was used as a control. Growth in the range of 2000 CFU per mL was obtained in the broth control by 24 hours, whereas the whole blood and buffy coat units appeared sterile for the first days of storage. After 1 week, a trace of bacteria and, after 4 weeks, massive growth were found in the WBC-containing units but not in the WBC-reduced units. The likely explanation is that the bacteria had been phagocytosed by the WBCs and were thereby hidden and not available for bacterial culture during the first phase of storage. When the WBCs spontaneously disintegrated, bacteria were released and multiplied in the blood units.(ABSTRACT TRUNCATED AT 250 WORDS)
Removal of buffy coat from the red cell preparations reduces the content of microaggregates and leukocytes and decreases transfusion side effects. However, buffy coat removal is difficult to standardize when performed manually. An automated method using a new type of plastic container system, the Optipac, and a new type of device, the Optipress, greatly improves blood component production. An initial hard centrifugation of the original whole blood provides the best results, leaving approximately 20 ml of plasma and 0.5 x 10(9) leukocytes per unit of red cells suspended in 100 ml of an additive solution (SAG-M). Improved quality and standardization of the products, reduced technician time and better working conditions for the technician are the major advantages as compared to traditional manual technique.
Blood collection and component preparation have been performed in integrally connected multiple plastic containers made with a new plastic. This polyvinylchloride (PVC) container plasticized with butyryl-n-trihexyl-citrate (BTHC) is a new material for blood storage; it contains no di(2-ethylhexyl)phthalate (DEHP). After removal of plasma and buffy coat, the red cells were suspended in saline-adenine-glucose-mannitol (SAGM) medium. After 42-day refrigerator storage, the total adenine nucleotide concentration remained the same as the initial concentration in the red cells, whereas ATP levels had decreased to 61 percent of the initial value. The 2,3 DPG concentration was 62 percent of normal on Day 7 and 21 percent on Day 14. Glucose consumption, lactate production, potassium leakage from red cells, and pH levels were similar to those found after storage in DEHP-plasticized containers under the same conditions. After 42 days, hemolysis levels were 0.56 +/- 0.21 percent and 0.42 +/- 0.17 percent in two series of units mixed weekly and 0.70 +/- 0.27 percent in units stored unmixed. Although even higher levels of hemolysis were observed in the units stored unmixed and used for 24-hour posttransfusion survival, the autologous red cell recovery results were excellent (83.2 +/- 5.1%, n = 8). BTHC-plasticized PVC is found to be a suitable material for 42-day storage of red cells in SAGM solution.
The possible beneficial role of white cells (WBCs) in donor blood has been investigated with respect to their capacity to remove bacteria. Preparations of buffy coat and whole blood, containing as well as reduced of WBCs, were inoculated with Staphylococcus epidermidis, S. aureus, Escherichia coli, Pseudomonas aeruginosa, and Propionibacterium species. Upon storage at room temperature, the presence of WBCs resulted in a reduction of the bacterial content. Units inoculated with S. epidermidis and E. coli were completely cleared of bacteria within 5 to 24 hours. On the other hand, S. aureus, after an initial reduction in number, started to multiply. In WBC-reduced units, the initial bacterial content remained unchanged for 5 hours, but the bacteria then exhibited vigorous growth within 48 hours in buffy coat and slower growth in whole blood. Propionibacterium sp. did not grow with or without WBCs. P. aeruginosa did not grow in buffy coat but showed a growth pattern similar to that of S. aureus in whole blood. The presence of WBCs in the donor blood during the first hours after collection thus seems to rid the blood of at least some species of bacteria. These results indicate that it would be favorable not to perform WBC reduction during blood collection and that several hours of contact can be needed to obtain sterility.
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Blood is normally collected into a combined anticoagulating and preserving medium. We performed a study to ascertain whether improvements could be made by separation of these two functions. Addition of saline-adenine-glucose solutions (40 to 100 ml per blood unit) to buffy-coat-poor red-cell concentrates allowed storage for as long as 35 days with 24-hour erythrocyte post-transfusion survival of 83 +/- 6.8 per cent (+/0 S.D.). Potassium leakage was lower, and in vitro hemolysis somewhat higher than that of whole blood. The microaggregate content after 21 days was 16 per cent of that in whole blood. In over-pressure transfusions the flow rate of red cells was the same with red-cell concentrates to which 80 to 100 ml of suspension medium had been added (hematocrit less than or equal to 60 per cent) as with whole blood. Removal of the buffy coat was essential to reduce hemolysis. We conclude that red cells can be successfully stored in a simple protein-poor medium.
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The kidney graft survival in 117 transplanted patients was investigated with reference to blood transfusions. No improvement of graft survival was observed in the transfused patients as compared to the patients who were not transfused or had got autologous blood only.
(1) Blood was stored in polyvinyl-chloride bags containing citrate-phosphate-dextrose (CPD) with adenine in a final concentration of 0.25 mM. (2) Red cell ATP was well maintained (greater than 70% of original) for 4 weeks in whole blood as well as in red cell concentrate (PCV 85 plus or minus 2%). After 5 weeks the ATP level was about 70% in whole blood and about 40% in red cell concentrate. (3) Red cell 2,3-diphosphoglycerate (DPG) was about 60% of the original after 2 weeks and about 30% after 3 weeks of storage when stored both as whole blood and as red cell concentrate. (4) The red cell 24-hour post-transfusion viability was about 80% after 4 weeks of storage both as whole blood and as red cell concentrate. After 5 weeks of storage the 24-hour viability was 78.7 plus or minus 3.5% in whole blood and 76.5 plus or minus 6.7% in red cell concentrate. (5) 820 patients received 3,238 units of CPD-adenine blood, and 761 patients serving as controls received 2,807 units of acid-citrate-dextrose (ACD) blood. The frequency of transfusion reactions was 3.5% for patients receiving CPD-adenine blood and 4.1% for the control group. (6) The maximum storage time was set at 5 weeks for the CPD-adenine blood and 3 weeks for the ACD blood. The longer preservation time decreased out-dating by at least 50%.
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Some technical improvements of the method to prepare platelet concentrates (PC) have been developed. The contamination of the PC with red blood cells (RBC) and white blood cells (WBC) is thereby kept at a low level. A majority of routinely prepared PC did not show any visible RBC contamination which means that they contained less than 0.4 X 10(9) RBC per PC unit. None of the tested examples contained more than 1 X 10(9) RBC per PC unit. The WBC contamination was less than 0.1 X 10(9) cells per PC unit in 75 per cent of tested examples and did not exceed 0.6 X 10(9) cells per PC unit. Routine platelet counting by thrombocounter applied on samples from all PRPs was found to be a reasonably simple way to make routine PC quality control. By sufficient supervision of the details of the procedure the main yield can be maintained at about 100 X 10(9) cells per unit. By follow-up of the weekly yield any deterioration of production efficiency can easily be detected.