Citrate Phosphate Dextrose Research Articles

Hematopoietic Progenitor Content of Fetal Cord Blood Collected Using Citrate-Phosphate-Dextrose: Influence of Holding Temperature and Delays

Storage of unseparated fetal cord blood collected into citrate-phosphate-dextrose (CPD) for 48 hours has been studied using progenitor cell assays [plasma clot technique with plasma-conditioned medium, erythropoietin (EPO), erythropoietin + IL-3 as growth factors]; collected fetal blood volume was low [61 +/- 4 ml, mean +/- 95% confidence interval (CI)]. The CFU-GM concentration in total cord blood was 2460 +/- 1476 CFU-GM/ml, comparable to normal bone marrow collected into medium RPMI + heparin (2350 +/- 1258); 2 of 12 samples contained fewer than 10(3) CFU-GM. Storage of unseparated cord blood CFU-GM in CPD was better at 4 degrees C (rapid decrease at day (D) 2 from 15 +/- 9 x 10(4) at D0; 14 +/- 11 x 10(4) at D1 to 6 +/- 4 x 10(4) at (D2) than at 22 degrees C (strong decrease since D1: 5 +/- 4 x 10(4) CFU-GM). The CFU-GEMM content remained steady at 4 degrees C: from 16 +/- 16 x 10(4) at D0 to 18 +/- 16 at D2. Cryopreservation and thawing of unfractionated cord blood at D0 did not affect the CFU-FM cloning rates. Whether mature progenitors are more affected than stem cells by storage in CPD remains to be determined.

Antimony electrode for the determination of the intramyocardial pH during open-heart surgery.

The development of an antiomony electrode (Sb) and a pH meter as a substitute for a glass electrode(G) to measure the intramyocardial pH is reported, and the results of their clinical application. The determination of the pH of CPD (citrate-phosphate-dextrose) blood by Sb showed small differences when compared to G (less than 0.1). Sb was also temperature sensitive. The temperature coefficient of Sb was determined in buffers by varying their temperatures, and an antimony-thermocouple electrode and a temperature-compensated pH meter were subsequently constructed. In CPD blood of varying temperatures Sb showed similar tendencies to G. The results of the application of the new apparatus to 28 patients were statistically no different from those obtained previously by G on 22 patients, either in baseline pH or its fall following aortic cross-clamping. It is concluded that although Sb is not as accurate as G, it is a reasonable alternative for the determination of myocardial acidosis during open-heart surgery.

Compatibility of Packed Erythrocytes and Ringer??s Lactate Solution

Packed erythrocytes are frequently reconstituted with crystalloid during rapid infusion. Dilution of whole blood with calcium containing solutions, such as Ringer's lactate has been cautioned against, citing possible clot formation because of chelation of the citrate anticoagulant. We studied the compatibility of Ringer's lactate solution and citrate phosphate dextrose (CPD)-preserved packed erythrocytes to evaluate the safety of using Ringer's lactate solution as a diluent in the emergency setting. Aliquots of CPD-preserved packed erythrocytes were diluted with either Ringer's lactate or normal saline solutions in ratios between 5:1 to 1:20 (packed erythrocyte to crystalloid), incubated at room temperature or 37 degrees centigrade and examined for clot formation at intervals up to two hours. Although clotting occurred at dilutions of 1:1 (packed erythrocytes to Ringer's lactate solution) and beyond, no clot formation occurred in the clinically relevant dilutions between 5:1 and 2:1. Thirty-two additional units of CPD-preserved packed erythrocytes were diluted to hematocrit values of 35, 45, 55 or 65 per cent and passed through a 170 micron filter. Flow rates of packed erythrocytes diluted with Ringer's lactate and normal saline solutions were compared. There was no difference in flow rates between packed erythrocytes diluted with Ringer's lactate compared with normal saline solutions. Ringer's lactate solution can be safely used as a packed erythrocyte diluent in patients requiring rapid blood transfusions.

Effect of Blood Transfusion on Acid Base, Glucose and Electrolyte Status in Very Low Birth Weight Infants

The effect of citrate phosphate dextrose (CPD) blood transfusion on the acid base, glucose and electrolyte status of VLBW infants was studied in infants aged 1-7, 8-20 and greater than 20 days; a 4th group composed of infants greater than 20 days subjected to a second transfusion was also studied. All the infants remained clinically well during and after the transfusion. Only the infants in the group 1-7 days experienced a significant fall in ionic Ca and PO2. No significant change in all the other parameters studied was observed. In the other groups of infants studied, the changes observed were not significant, and became less with increasing age and repeat transfusion. VLBW infants apparently tolerate CPD blood transfusion well.

赤血球の液状の保存温度は何度まで下げられるか？

The present study was designed to find the lowest acceptable temperature for liquid preservation of red blood cells. We stored red cells at 5°C, 2.5°C, 0°C, -2°C and -5°C for 6 weeks in citrate-phosphate-dextrose (CPD) solution. Supernatant hemoglobin concentration was maintained at the low level at 0°C throughout the storage, but showed a significant increase at a temperature below -2°C. A temperature of 0°C was the most effective in the maintenance of the adenosine triphosphate (ATP) level, morphology score, and erythrocyte deformability, indicating that the red cells stored at 0°C were the most viable. The 2, 3-diphosphoglycerate (2, 3-DPG) level during the storage increased in the order of 5°C<0°C<-2°C, -5°C. These data suggest that the temperature for red cell preservation can be lowered to 0°C. Judging from the ATP level and morphology score, the storage period of red blood cells could be extended up to about 5 weeks even in CPD solution if the red cells are stored at 0°C.

Zinc Ions Inhibit Factor I‐Mediated Release of CR1‐Bound Immune Complexes and Degradation of Cell‐Bound Complement Factors C3b and C4b

ZnCl2 exerted a dose-dependent inhibition of citrate-phosphate-dextrose (CPD) plasma-induced release of 125I-labelled BSA-anti-BSA immune complexes (IC) bound to complement receptor type 1 (CR1, CD35) in human whole blood. Maximal inhibition was observed at 10 mM of ZnCl2. Furthermore, the release of IC bound to erythrocyte (E)-CR1 by purified factor I, factor I-deficient serum plus purified factor I, or normal human serum was reduced by approximately 90%, 64%, and 52%, respectively, in the presence of 10 mM ZnCl2. The effect of ZnCl2 on factor I-mediated degradation of cell-bound C3b/C4b was also investigated employing CPD blood or E from a factor I-deficient donor. These cells expressed covalently bound C3b and C4b as demonstrated by a simple agglutination technique. Upon incubation of CPD whole blood with purified factor I, or of E with purified factor I or normal CPD plasma, the C-fragments were cleaved and the cells were no longer agglutinated by antibodies to C3c and C4c. The presence of ZnCl2 prevented this factor I-mediated degradation of C3b and C4b, as evidenced by the unaffected agglutination of the cells by the antibodies. We conclude that ZnCl2 inhibited factor I activity since: (1) release of complement-preopsonized IC from E-CR1 by purified factor I was markedly inhibited (90%) in the presence of ZnCl2, (2) preincubation of the cells with ZnCl2 caused only a moderate inhibition (32-38%) of the IC release, and (3) degradation by purified factor I of covalently cell-bound C3b and C4b was abrogated in the presence of 10 mM ZnCl2.

Effect of Serum Precoat on Complement Activation in Membrane Oxygenators: An In Vitro Study

Previous studies have shown that extracorporeal circulation devices can activate the complement cascade via the alternative pathway. Anaphylatoxins (C3a and C5a) generated by complement activation may lead to pulmonary leukocyte sequestration, pulmonary edema, granulocytopenia and microvascular lung damage. This is of particular concern in patients on cardiopulmonary bypass. The purpose of this study was to examine the impact on complement activation of precoating membrane oxygenators with serum. A pool of recalcified human serum was prepared from fresh frozen plasma stored in citrate phosphate dextrose. SciMed membrane oxygenators (N=4) were primed with 200 ml aliquots of pooled serum which were recirculated through the oxygenators for 180 minutes at 37° C. The primary recirculation serum C3a levels (mean ± SD) measured by radioimmunoassay techniques at 0, 60, 120, and 180 minutes were 122.25 ± 26.22, 430.25 ± 164.39, 456.0 ± 154.16 and 577.5 ± 163.07 ng/ml respectively. Each oxygenator was then flushed with saline and reprimed with a fresh 200 ml aliquot of pooled serum which was recirculated for 180 minutes at 37° C. The secondary recirculation serum C3a levels (mean ± SD) measured at 0, 60, 120, and 180 minutes were 130.75 ± 15.67, 213.25 ± 58.24, 283.75 ± 27.24 and 301.75 ± 19.18 ng/ml respectively. Serum C3a levels were higher in the primary serum than the secondary serum in all oxygenators. These results suggest that preconditioning the oxygenator may be a way to moderate complement activation during cardiopulmonary bypass.

４連バッグシステムを用いたＢＵＦＦＹ ＣＯＡＴ除去濃厚赤血球の調製

Whole blood (400ml) was collected in citrate-phosphate dextrose in quodruple bag system. After centrifugation for 6min at 2, 200rpm, blood was separated into platelet rich plasma, huffy coat (approximately 45ml), and concentrated red cells. The platelet rich plasma was separated into platelet concentrates and plasma. The buffy coat contained approximately 98% lymphocytes, 70% granulocytes and 12% red cells of the original value. The leukocyte contents of huffy-coat-depleted concentrated red cells were 50±28×107per unit, in particular, lymphocytes decreased to 1.4±0.5×107per unit, because of the most lymphocytes were removed with buffy coat. This system seems to be as safe and effective for preparation of leukocytes poor red cells, and can be operated without machines.

Decrease in Hepatic Mitochondrial Redox State by Massive Infusion of Citrate-Phosphate-Dextrose Solution in Jaundiced Rabbits

To clarify the mechanism of metabolic derangement by massive blood transfusion to the damaged liver, the changes in the hepatic mitochondrial redox state, as reflected in arterial ketone body ratio (acetoacetate/3-hydroxybutyrate), were studied in jaundiced rabbits by infusion of massive citrate-phosphate-dextrose (CPD) solution. The jaundiced rabbits received common bile duct ligation (BDL group), and the sham group had a simple exploration of the bile duct. CPD solution was infused for 3 h at a rate of 9 ml/kg/h in each group. As metabolic parameters, blood gas, pyruvate, lactate, citrate and ketone body ratio were analyzed in the arterial blood. During the course, arterial ketone body ratio decreased significantly (p less than 0.01) in the BDL group with severe metabolic acidosis, while it was maintained at high value in the sham group with metabolic alkalosis. Organic acids were more highly accumulated in the BDL group than in the sham group. These results suggested a hepatodepressant effect of massive blood transfusion, especially in the damaged liver.

Infectivity of cultured Plasmodium falciparum gametocytes to mosquitoes.

Various factors that may influence routine and high levels of mosquito infection with cultured Plasmodium falciparum gametocytes are considered in this paper. One of the most important is the choice of an appropriate isolate, with facilities for cryopreservation and a good technique for initiation of cultures. The use of automated culture systems with strict adherence to detail and routine has eliminated much of the variability. The quality of the serum used for the culture of gametocytes and inclusion in the feed material for mosquitoes is of the highest importance. Blood collection for culture purposes must preferably involve alcohol as an antiseptic for cleaning donor skin or suitable receptacles. Mosquito blood meals should not include plasma with citrate phosphate dextrose or sera collected in microtainer tubes or from volunteers on proguanil-chloroquine prophylaxis. Sera of individuals on chloroquine alone do not influence transmission. Haematocrits of from 5 to 10% permit the culture of equally infective gametocytes. It was impossible to predict the outcome of an infection in mosquitoes based on the number of female gametocytes or gametes. Within any experiment, the oocyst load initially increased, followed by a decline with progressively lower numbers of gametocytes accompanied by a progressive increase in the efficiency of transmission. Some of the variability of mosquito infection within an experiment was due to individual differences in the speed of blood digestion of the mosquitoes. A new membrane feeder is described with three different sizes to accommodate a variety of goals.(ABSTRACT TRUNCATED AT 250 WORDS)

Compatibility of ADSOL-stored red cells with intravenous solutions

Five percent dextrose in water (D5W) and lactated Ringer's (RL) are two intravenous solutions that are incompatible with citrate phosphate dextrose (CPD) anticoagulated RBCs. Hemolysis, agglutination, and clotting can occur when RBCs are mixed with, infused together with, or transfused in the same line following administration of these fluids. Although minimal plasma remains in units of RBCs containing adenine-saline-dextrose solution (ADSOL; Fenwal Laboratories, Deerfield, IL), clots are seen immediately after contact is made with RL. Clumping and hemolysis are seen when ADSOL-stored RBCs or sallne-washed RBCs are mixed with D5W. This study reinforces the unsafe practice of mixing RBCs with certain intravenous solutions.

Serial Determinations of PF4 and βTG: Comparisons Between Multiple Venipunctures vs a Catheter Infusion System

Platelet factor 4 (PF4) and beta thromboglobulin (beta TG) are platelet-specific proteins which are released upon platelet aggregation and which can be accurately measured by radioimmunoassay. We devised a catheter-infusion system that enables serial determinations of these proteins. In 20 subjects (10 healthy volunteers and 10 patients with stable coronary artery disease), we compared samples collected by individual venipunctures with those simultaneously obtained by means of a simple catheter-infusion system. At least 5 samples were obtained over a period of time which was as long as 60 min, and at least 30 min. Subjects with stable coronary artery disease were selected so that they would be expected to have stable and normal PF4 and beta TG levels. Thus, elevations of either PF4 or beta TG would represent artifacts secondary to sampling technique. Analysis of the results demonstrated that the catheter-infusion system was equivalent to individual venipunctures for determination of PF4 and beta TG. 16.8% of samples obtained via the catheter and 17.2% of those obtained by individual venipunctures were spuriously elevated. A second series of studies were performed to refine the technique further by examining the impact of infusion rate and the addition of citrate phosphate dextrose (CPD) to the infusate. Ten additional subjects had catheter systems utilized in both arms simultaneously. The addition of CPD resulted in significantly less abnormal values at slower infusion rates (1 and 2.5 cc/min). At 5 cc/min D5/w or saline alone are suitable. These investigations confirm that this simple catheter system is equivalent to individual venipunctures for determination of PF4 and beta TG while avoiding patient discomfort.(ABSTRACT TRUNCATED AT 250 WORDS)

Preparation of leukocyte-poor platelet concentrates from buffy coats. II. Lack of effect on storage of different plastics.

Six citrate phosphate dextrose (CPD)-saline adenine glucose mannitol (SAG M) quadruple systems were evaluated for the preparation and storage of leukocyte-poor platelet concentrates (PC) from buffy coats. The platelet storage bags examined were manufactured from normal polyvinylchloride (PVC) or special-type plastics. Biotest supplied PVC 76 (n = 14) and PVC 763 (n = 16) NPBI supplied PSV 3277 (n = 15) and DPL-110 (n = 14) and Terumo supplied Teruflex (n = 18) and molded Teruflex (n = 14). The PC were stored for 7 days at 22 degrees C on a horizontally shaking platform. Cell counts, pH, PO2, PCO2, morphology score and swirling patterns were monitored at 5, 72, 120 and 168 h. The plasma volumes averaged 63 ml and ranged from 39 to 81 ml, the overall mean +/- SD platelet concentration was 0.89 +/- 0.33 X 10(9)/ml. None of the PC had a leukoyte count higher than 10 X 10(6) per unit. After storage for 168 h, the pH ranged from 6.56 to 7.40 for all brands. The PO2 remained stable and even rose significantly (p less than 0.05) during storage in the NPBI PSV 3277 and Terumo molded Teruflex bags. The PCO2 decreased equally in all bags. Morphology scores were well maintained in 98% of all PC for up to 120 h, and in 83% at 168 h. A swirling pattern score of 2.5 or greater predicted with a specificity of 100% a good morphology score in the PC.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of oxalate and malonate on red cell metabolism

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

Survival of red cells stored for 21 and 35 days in a non-di-(2-ethylhexyl)phthalate plastic container.

Whole blood and red cells were stored using citrate-phosphate-dextrose (CPD) and citrate-phosphate-dextrose-adenine (CPDA-1) anticoagulants in polyvinylchloride bags made flexible with di-(2-ethylhexyl)phthalate (DEHP) or tri-(2-ethylhexyl)trimellitate (TOTM) plasticizers. After storage the posttransfusion viability of these cells was tested in autologous donors. Cells stored in TOTM-plasticized film had a survival rate less than 75% when stored for 35 days, while other systems had a survival greater than this. When compared with the red cells stored in CPD-DEHP-plasticized film, the viability of whole blood and red cells stored in CPDA-TOTM showed a statistically significant decrease (p = less than 0.01). Therefore, red cell storage in TOTM-plasticized PVC with current anticoagulant should be limited to 21 days.

Effect of Oxalate and Malonate on Red Cell Metabolism

The addition of oxalate to blood stored in Citrate-phosphate-dextrose (CPD) produces a marked improvement in 2,3-diphosphoglycerate (2,3-DPG) preservation; an increase in 2,3-DPG levels can also be documented in short-term incubation studies. Oxalate is a potent in vitro inhibitor of red cell lactate dehydrogenase, monophosphoglycerate mutase, and pyruvate kinase (PK). In the presence of fructose 1,6-diphosphate the latter inhibitory effect is competitive with phospho(enol)pyruvate (PEP). Determination of the levels of intermediate compounds in red cells incubated with oxalate suggest the presence of inhibition at the PK step, indicating that this is the site of oxalate action. Apparent inhibition at the glyceraldehyde phosphate dehydrogenase step is apparently due to an increase in the NADH/NAD ratio. Oxalate had no effect on the in vivo viability of rabbit red cells stored in CPD preservatives for 21 days. Greater understanding of the toxicity of oxalate is required before it can be considered suitable as a component of preservative media, but appreciation of the mechanism by which it affects 2,3-DPG levels may be important in design of other blood additives. Malonate, the 3-carbon dicarboxylic acid analogue of oxalate late did not inhibit pyruvate kinase nor affect 2,3-DPG levels.

Storage-induced changes in human newborn red cells.

Fetal red cells are well suited for intrauterine life; however, little is known about their response to postnatal environments. The purpose of this work was to investigate the metabolic and membrane changes affecting newborn red cells during their exposure to storage in citrate-phosphate-dextrose (CPD) and citrate-phosphate-dextrose-adenine (CPDA-1). The findings suggest that newborn red cells are affected more by storage than are adult cells. These accelerated storage changes in the red cells of newborns may be related indirectly to the rapid adenosine triphosphate (ATP) decline. As is the case with adult red cells, fetal cells withstand storage in CPDA-1 better than in CPD. The storage lesion in these cells was partly reversible, as in adult cells, by incubation with adenosine. It was therefore concluded that newborn red cells obtained from placentas and stored for several weeks in CPD or CPDA-1 media or other media that improve the metabolic profile of these cells may be acceptable for transfusion.

The use of nystatin to restore the flow properties of time-expired stored erythrocytes.

Human erythrocytes stored for more than 21 days in citrate phosphate dextrose with adenine (CPDA-1) exhibited a marked reduction in volume following incubation for 24 h at 37 degrees C in fresh autologous plasma. This incubation apparently mimics the effects of reinfusion of the stored cells in vivo. These shrunken stored cells have a decreased filterability as measured by their increased transit times through a 5-microns diameter Nuclepore filter. The polyene antifungal agent nystatin was used to reinflate these shrunken cells with different concentrations of potassium ions. A concentration of 90 mM potassium chloride was found to reinflate the shrunken stored cells so that their mean cell volume, mean corpuscular haemoglobin concentration and average densities were the same as those of fresh cells. This reinflation also restored the filterability of the shrunken stored cells so as to be similar to that of fresh cells.

The role of mean corpuscular haemoglobin concentration in limiting the storage life of human blood.

Human erythrocytes stored for more than 21 days in citrate phosphate dextrose with adenine (CPDA-1) at +4 degrees C had a decreased cell volume, an increased cell density, an elevated mean corpuscular haemoglobin concentration (MCHC) and hence an elevated internal viscosity. These changes are normally masked by the reversible cell swelling which accompanies storage in CPDA-1. Incubation of stored cells for 24 h at 37 degrees C in fresh autologous plasma mimics the effects of reinfusion and causes the artificially swollen cells to shrink to their true volume. Thus, incubated cells which had been stored in CPDA-1 for more than 21 days exhibited a decreased filterability through Nuclepore membranes in vitro, which is apparently correlated with a decreased survival time in vivo.

The use of banked autologous blood in patients undergoing surgery for spinal deformity

The cases of fifty-two patients who underwent sixty elective spinal fusions for spinal deformity were studied to evaluate the efficacy of the use of banked autologous blood to replace operative loss of blood. The patients ranged in age from ten to forty-nine years. Each patient began to take 325 milligrams of ferrous sulphate, three times a day, as soon as surgery was scheduled, and was evaluated weekly at the Shepeard Community Blood Bank. If a patient's hemoglobin level was more than eleven milligrams per 100 milliliters, either a whole unit of blood or a half-unit was drawn at each visit. An average of 3.3 units of blood (range, 1.5 to 6.0 units) was obtained and was stored for as long as forty-two days. Either citrate phosphate dextrose with adenine (CPDA-1) or adenine, dextrose, and mannitol (ADSOL) was used as a preservative. In 85 per cent of the procedures only autologous blood was required for transfusion. This method proved to be simple, safe, and very well accepted.