24-hour Posttransfusion Survival Research Articles

Early and Late-Phase 24h Responses of Stored Red Blood Cells to Recipient-Mimicking Conditions.

The 24-hour (24 h) post-transfusion survival of donor red blood cells (RBCs) is an important marker of transfusion efficacy. Nonetheless, within that period, donated RBCs may encounter challenges able to evoke rapid stress-responses. The aim of the present study was to assess the effect of exposure to plasma and body temperature upon stored RBCs under recipient-mimicking conditions in vitro from the first hours “post-transfusion” up to 24 h. For this purpose, packed RBCs from seven leukoreduced CPD/SAGM units were reconstituted with plasma of twenty-seven healthy individuals and incubated for 24 h at 37oC. Three units were additionally used to examine stress-responses in 3-hour intervals post mixing with plasma (n = 5) until 24 h. All experiments were performed in shortly-, medium-, and long-stored RBCs. Hemolysis, redox, morphology, membrane protein binding and vesiculation parameters were assessed. Even though spontaneous hemolysis was minimal post-reconstitution, it presented a time-dependent increase. A similar time-course profile was evident for the concentration of procoagulant extracellular vesicles and the osmotic fragility (shortly-stored RBCs). On the contrary, mechanical fragility and reactive oxygen species accumulation were characterized by increases in medium-stored RBCs, evident even from the first hours in the recipient-mimicking environment. Finally, exposure to plasma resulted in rapid improvement of morphology, especially in medium-stored RBCs. Overall, some RBC properties vary significantly during the first 24 h post-mixing, at levels different from both the storage ones and the standard end-of-24 h. Such findings may be useful for understanding the performance of RBCs and their possible clinical effects −especially on susceptible recipients− during the first hours post-transfusion.

Commentary on the Sterility of Blood Products and the Measurement of the 24-Hour Posttransfusion Survival of Preserved RBC using 51Cr Single Label and 51Cr Double Label Procedures

This commentary is written to respond to recent publications on the storage of whole blood at room temperature for 24 hours prior to leukoreduction and storage of plasma at less than -18 C for one year and red blood cells stored at 4 C in Additive Solution 7 for 42 and 56 days and blood stored at room temperature for less than 2 hours and 6 to 8 hours prior to storage of RBC in the Additive Solution 7 for 42 and 56 days. The authors of these 3 papers provided no data on the sterility of these blood products. In addition, the 24 hour posttransfusion survival values of autologous red blood cell stored at 4 C in the additive solution AS-7 for 42 and 56 days were performed using a 51Cr single label procedure. Introduction

Commentary on the Sterility of Blood Products and the Measurement of the 24-Hour Posttransfusion Survival of Preserved RBC using 51Cr Single Label and 51Cr Double Label Procedures

This commentary is written to respond to recent publications on the storage of whole blood aroom temperature for 24 hours prior to leukoreduction and storage of plasma at less than -18 C for one year and red blood cells stored at 4 C in Additive Solution 7 for 42 and 56 days and blood stored at room temperature for less than 2 hours and 6 to 8 hours prior to storage of RBC in the Additive Solution 7 for 42 and 56 days. The authors of these 3 papers provided no data on the sterility of these blood products. In addition, the 24 hour posttransfusion survival values of autologous red blood cell stored at 4 C in the additive solution AS-7 for 42 and 56 days were performed using a 51Cr single label procedure. Introduction

Volume of RBCs, 24- and 48-hour posttransfusion survivals, and the lifespan of (51)Cr and biotin-X-N-hydroxysuccinimide (NHS)-labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on (51)Cr and biotin-X-NHS elution in vivo.

The RBC injury that occurs during collection of the first few milliliters of blood into the pH 5.0 ACD (NIH, Formula A) is referred to as the lesion of collection. The RBC injury was evaluated by labeling the ACD RBCs with (51)Cr and measuring the 24-hour posttransfusion survival. The effect of the acidification of ACD blood on the in vivo elution of (51)Cr from the RBC has not been reported. Baboon blood was collected in heparin and in ACD and CP2D at different ratios of blood to anticoagulant. ACD blood with a pH of 5.7 to 6.9 was labeled with (51)Cr. Heparinized blood with a pH of 7.4 was labeled with biotin-X-N-hydroxysuccinimide (NHS). ACD blood with a pH of 5.9 was labeled with both (51)Cr and biotin-X-NHS. The RBC volumes, 24- and 48-hour posttransfusion survivals, and lifespans (T50) were measured. The RBC volume of ACD blood with a pH ranging from 5.7 to 6.9 was not affected by (51)Cr labeling. (51)Cr-labeled ACD blood with a pH of 6.9 had an RBC volume that was significantly greater than that seen in heparinized blood with a pH of 7.4 labeled with biotin-X-NHS. In vivo elution of (51)Cr from the RBCs prepared from the ACD blood with a pH of 5.7 to 6.9 and in vivo elution of biotin-X-NHS from RBCs prepared from ACD blood with a pH of 5.9 were associated with reductions in 24- and 48-hour posttransfusion survivals and T50. The anticoagulant and the pH of the medium in which the RBCs were labeled with (51)Cr or biotin-X-NHS affected in vivo elution of the label from the RBCs and may reduce posttransfusion survival values.

Survival, function, and hemolysis of shed red blood cells processed as nonwashed blood and washed red blood cells

Background. Shed nonwashed blood and shed washed red blood cells (RBC) are being used as alternatives to allogeneic liquid-preserved RBC for patients during thoracic and cardiovascular surgical procedures. Methods. Mongrel dogs were bled a volume of blood into the abdominal cavity and the shed blood was reinfused as nonwashed blood or washed RBC. The 51Cr RBC volumes were measured before, immediately after, and 24 hours after the exchange transfusion to assess the recovery of the shed RBC and the 24-hour posttransfusion survival. Compatible dogs were given allogeneic transfusions of 51Cr-labeled nonwashed blood and washed RBC, and 24-hour posttransfusion survival and half-life were measured. Results. Immediately after the 100% exchange transfusion, the recovery value was 62% for the nonwashed shed blood and 82% for the washed RBC. Both the nonwashed blood and the washed RBC had 24-hour posttransfusion survival values of 90% and normal oxygen transport function after the exchange transfusion. Compatible allogeneic 51Cr-labeled nonwashed blood and washed RBC had normal 24-hour posttranfusion survival and 51Cr half-life values. Conclusions. The survival, function, and hemolysis of shed nonwashed blood and shed washed RBC were similar to fresh blood in the dog that underwent a 100% exchange transfusion.

Survival and function of baboon RBCs released from clotted blood and washed before autologous transfusion.

One alternative to an allogeneic transfusion is the salvaging of the patient's own shed blood. In this study, baboon blood was allowed to clot and the RBCs that were released from the clotted blood lysed with and without urokinase were washed before autologous transfusion. Forty-four studies were done in 13 baboons (Papio cynocephalus or Papio anubis) over a 3-year period. In 24 studies, a 50-mL volume of blood was collected without an anticoagulant and stored at 22 degrees C for as long as 72 hours before washing and autologous transfusion. In 20 other studies, a 50-mL volume of blood was collected without an anticoagulant and allowed to clot for 30 to 60 minutes. Urokinase, ranging from 2,500 to 10,000 units per mL, was added, and the blood was stored at 22 degrees C for 24 hours before washing and autologous transfusion. RBCs that were stored at 22 degrees C without urokinase for 24 hours exhibited an in vitro recovery value of 45 percent, a (51)Cr 24-hour posttransfusion survival of 86 percent, and an index of therapeutic effectiveness of 39 percent. The (51)Cr T(50) value was normal at 14 days, and RBC oxygen-transport function was slightly reduced. RBCs that were stored at 22 degrees C for 24 hours with 10,000 units per mL of urokinase exhibited an in vitro recovery value of 89 percent, a (51)Cr 24-hour posttransfusion survival value of 86 percent, and an index of therapeutic effectiveness of 76 percent. The (51)Cr T(50) value was normal at 14 days, and the RBC oxygen-transport function was only slightly reduced. Autologous baboon RBCs isolated from clotted blood treated or not treated with urokinase and washed before transfusion have excellent survival and normal or only slightly reduced oxygen-transport function.

In vivo survival of apheresis RBCs, frozen with 40-percent (wt/vol) glycerol, deglycerolized in the ACP 215, and stored at 4 degrees C in AS-3 for up to 21 days.

The FDA has approved the storage of frozen RBCs at -80 degrees C for 10 years and the postwash storage at 4 degrees C for no more than 24 hours. The 4 degrees C postwash storage period is limited to 24 hours, because the current deglycerolization systems are functionally open systems. Two units of RBCs were collected from each of 13 healthy male volunteers. The RBCs were collected in CP2D by the FDA-approved protocol for an automated apheresis device (MCS, LN8150, Haemonetics) and were stored at 4 degrees C in AS-3 for 6 days. Using a single disposable glycerolization set in an automated, functionally closed system (ACP 215, Haemonetics) each unit was transferred to a 1000-mL PVC plastic bag and glycerolized to a concentration of 40-percent (wt/vol) glycerol and frozen at -80 degrees C. A single disposable deglycerolization set in the ACP 215 was used to deglycerolize the 2 units from the same donor. The deglycerolized RBCs were stored at 4 degrees C in AS-3 for as long as 21 days. The mean +/- SD freeze-thaw-wash recovery value was 89.4 +/- 3 percent. The residual hemolysis in the RBCs stored at 4 degrees C in AS-3 for 21 days after deglycerolization was 0.9 +/- 0.2 percent, and the units were negative for both aerobic and anaerobic bacteria. The mean Nageotte WBC count was 9 x 10(6) per unit. When the deglycerolized RBCs were given as an autologous transfusion after storage at 4 degrees C in AS-3 for the 7- to 18-day period, the mean +/- SD 24-hour posttransfusion survival was 77 +/- 7 percent, and the index of therapeutic effectiveness was 69 +/- 8 percent. Two units of human RBCs collected from a single donor by apheresis in the MCS using an LN8150 set can be glycerolized sequentially with a single disposable set and deglycerolized sequentially with another single disposable set in the ACP 215. The previously frozen RBCs stored in AS-3 for 7 to 18 days at 4 degrees C had acceptable hemolysis and an acceptable mean 24-hour posttransfusion survival value and index of therapeutic effectiveness.

In vitro and in vivo measurements of gamma-radiated, frozen, glycerolized RBCs.

Transfusion-associated GVHD results from the presence of viable lymphocytes in transfused allogeneic blood components. Viable immunocompetent lymphocytes have been detected in RBCs that were frozen with glycerol and washed before transfusion. The study reported here assessed the effect of irradiation on human RBCs frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. In vitro and in vivo testing was done on human RBCs that were frozen with 40-percent (wt/vol) glycerol at -80 degrees C, with some units exposed to 2500 cGy of gamma radiation and others not irradiated, and that, after thawing and washing, were stored in a sodium chloride-glucose solution at 4 degrees C for 3 days before autologous transfusion. The glycerol-frozen RBCs treated with 2500 cGy before deglycerolization had a mean freeze-thaw-wash recovery of 87 percent and a mean 24-hour posttransfusion survival of 86 percent after storage for 3 days at 4 degrees C in a 0.9-percent NaCl and 0.2-percent glucose solution. For the nonirradiated units, the mean freeze-thaw-wash recovery was 85 percent and the mean 24-hour posttransfusion survival was 83 percent. These data show similar, acceptable results for RBCs frozen with 40-percent (wt/vol) glycerol at -80 degrees C and treated in the frozen state with 2500 cGy of gamma radiation and for RBCs that were not irradiated, all of which were washed and then stored in a sodium chloride-glucose solution for 3 days before autologous transfusion.

Posttransfusion survival (24-hour) and hemolysis of previously frozen, deglycerolized RBCs after storage at 4 degrees C for up to 14 days in sodium chloride alone or sodium chloride supplemented with additive solutions.

Previously frozen human RBCs currently are glycerolized and deglycerolized by the use of open systems that limit storage of the deglycerolized RBCs at 4 degrees C to only 24 hours. Healthy male volunteers who met AABB requirements for blood donors (n = 38) were studied. A volume of 450 mL of blood was collected into CPDA-1. The RBC concentrates were stored at 4 degrees C for 3 to 6 days before being frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. The RBCs were deglycerolized, resuspended in 0.9-percent sodium chloride and 0.2-percent glucose (SG) solution or SG solution supplemented with AS-1, AS-3, or AS-5, and stored in the resuspension medium at 4 degrees C for 14 days. The mean +/- SD freeze-thaw-wash process recovery was 90.0 +/- 4.0 percent for all 38 units. The mean 24-hour posttransfusion survival value was 79 percent for deglycerolized RBC stored at 4 degrees C for 7 days in SG alone, SG plus AS-3, or SG plus AS-5. Deglycerolized RBC that were stored at 4 C for 14 days in SG supplemented with AS-1, AS-3, or AS-5 had a mean 24-hour posttransfusion survival of 74 percent. After 7 days of storage of deglycerolized RBCs in SG alone, the mean hemolysis was 3. 7 percent. After 14 days of storage of deglycerolized RBCs in SG supplemented with AS-1, AS-3, or AS-5, the mean hemolysis was 2.5 percent. The levels of hemolysis did not correlate with the 24-hour posttransfusion survival values.

Half-strength citrate CPD combined with a new additive solution for improved storage of red blood cells suitable for clinical use.

Currently used systems for red blood cell (RBC) collection and storage for transfusion have the disadvantage that the RBC 2,3-bisphosphoglycerate (BPG) concentration is depleted within two weeks of storage, resulting in a left-shift of the oxygen dissociation curve and a temporarily impaired capacity to deliver oxygen. We have studied the effects on red cell metabolism, morphology and in vivo recovery of 49-day storage of RBC, with collection in half-strength citrate CPD (0.5CPD) and storage in an additive solution containing citrate, adenine, mannitol, phosphate and glucose (RAS2). Traditional CPD-SAGM was used for comparison. Component preparation was performed after an initial holding period of the whole blood at ambient temperature for 8 h. The BPG concentration in 0.5CPD-RAS2 RBC was 0.633 +/- 0.120 mol (mol Hb)-1 as compared to 0.454 +/- 0.138 mol (mol Hb)-1 in CPD-SAGM RBC which implied a decrease to 67 and 48% of normal concentration, respectively. The mean RBC BPG concentration was maintained at the initial level for 28 days in the new system but decreased to very low levels within 14 days in the controls. The total adenine nucleotides were well maintained in both systems, adenosine triphosphate slightly better in the new system. Hemolysis after 49 days was 0.35 +/- 0.21% in the new system and 0.72 +/- 0.25% in the controls (p < 0.001). The morphology was better maintained in the new system (p < 0.001). The 24-hour posttransfusion survival of 49-day stored RBC was 78.9 +/- 7.1%. The membrane leakage of sodium and potassium was not significantly different in the two systems.(ABSTRACT TRUNCATED AT 250 WORDS)

Half-Strength Citrate CPD Combined with a New Additive Solution for Improved Storage of Red Blood Cells Suitable for Clinical Use

Currently used systems for red blood cell (RBC) collection and storage for transfusion have the disadvantage that the RBC 2,3-bisphosphoglycerate (BPG) concentration is depleted within two weeks of storage, resulting in a left-shift of the oxygen dissociation curve and a temporarily impaired capacity to deliver oxygen. We have studied the effects on red cell metabolism, morphology and in vivo recovery of 49-day storage of RBC, with collection in half-strength citrate CPD (0.5CPD) and storage in an additive solution containing citrate, adenine, mannitol, phosphate and glucose (RAS2). Traditional CPD-SAGM was used for comparison. Component preparation was performed after an initial holding period of the whole blood at ambient temperature for 8h. The BPG concentration in 0.5CPD-RAS2 RBC was 0.633±0.120 mol (mol Hb)^-1 as compared to 0.454±0.138 mol (mol Hb)^-1 in CPD-SAGM RBC which implied a decrease to 67 and 48% of normal concentration, respectively. The mean RBC BPG concentration was maintained at the initial level for 28 days in the new system but decreased to very low levels within 14 days in the controls. The total adenine nucleotides were well maintained in both systems, adenosine triphosphate slightly better in the new system. Hemolysis after 49 days was 0.35±0.21% in the new system and 0.72±0.25% in the controls (p<0.001). The morphology was better maintained in the new system (p<0.001). The 24-hour posttransfusion survival of 49-day stored RBC was 78.9±7.1%. The membrane leakage of sodium and potassium was not significantly different in the two systems. The concentration of inorganic phosphate (Pi) in the extracellular fluid of 0.5 CPD-RAS2 RBC was initially 7.1±0.8 mmol l^-1. It decreased to approximately 5.5 mmol l^-1 during the first 2 weeks of storage and then slowly increased to 9.4±0.9 mmol l^-1 on day 49. Pi in the CPD-SAGM controls was low initially but rose to 6.2±0.4 mmol l^-1 at the end of storage. The new system offers clearcut advantages as compared to traditional systems and does not seem to raise any serious clinical objections against its use even in massive transfusions.

Suitable survival and half‐life of red cells after frozen storage in excess of 10 years

To examine the appropriateness of the Food and Drug Administration's 10-year storage time for previously frozen red cells, 24-hour posttransfusion survival studies were performed, and the half-life of 3 units of autologous red cells that had been stored for 13.5, 14, and 17 years, respectively, was measured. The units had acceptable freeze-thaw-wash recovery (83.3-91.4%). When a 51Cr label was used for the previously frozen red cells and a simultaneous 52Cr label for freshly drawn autologous red cells was used as a comparison, it was seen that the previously frozen cells had normal 24-hour posttransfusion survival (75.1-88.4%) as well as normal half-life (23-33.7 days). These findings support further extension of the maximum allowable storage time for previously frozen red cells.

Storage of saline‐adenine‐glucose‐mannitol‐suspended red cells in a new plastic container: polyvinylchloride plasticized with butyryl‐n‐ trihexyl‐citrate

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 in vivo survival of human red cells with low oxygen affinity prepared by the osmotic pulse method of inositol hexaphosphate incorporation

The purpose of this study was to determine the posttransfusion survival of low-affinity red cells (RBCs) containing the hemoglobin modifier, inositol hexaphosphate (IHP). An osmotic pulse technique was used to incorporate IHP, a compound that normally does not cross the RBC membrane. Two groups of six volunteers were studied. RBCs were treated under conditions that resulted in a relatively large increase in mean corpuscular volume (delta MCV = 11.8 +/- 4.0, 1 SD) in Group I, but a relatively small increase (delta MCV = 5.5 +/- 1.9) in Group II. The groups had a similar decrease in mean corpuscular hemoglobin (delta MCH = -3.8 +/- 0.9 for Group I and -3.1 +/- 1.1 for Group II). The 24-hour posttransfusion RBC survival, measured with 3 mL of 51Cr-labeled autologous RBCs, correlated with the induced change in oxygen affinity, and larger shifts were associated with poorer survival. An acceptable 75-percent 24-hour posttransfusion survival was achieved for p50 increases up to approximately 15 torr. Cells that survived 24 hours appeared to have a normal lifespan. This study is the first demonstration of adequate survival for human RBCs with a potentially long-lasting decrease in oxygen affinity.

The safety and therapeutic effectiveness of human red cells stored at -80 degrees C for as long as 21 years.

Human red cells frozen by various methods have been stored in the frozen state at -80 degrees C for as long as 21 years. This report discusses: red cells frozen with 42 percent weight per volume (wt/vol) glycerol in an ionic medium in a polyvinylchloride (PVC) plastic bag using the Cohn method; red cells frozen with 45 percent wt/vol glycerol in a low ionic medium in a PVC plastic bag using the Huggins method; red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a polyolefin plastic bag using the Meryman-Hornblower method; and red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a standard 600-ml or an elongated 800-ml PVC plastic primary collection bag with an adapter port using the Naval Blood Research Laboratory (NBRL) method. After frozen storage for as long as 21 years by the four methods described above, the thawed red cells were deglycerolized with 50 to 150 ml of 12 percent sodium chloride and 1.5 to 2.0 l of sodium chloride-glucose or sodium chloride-glucose-phosphate solution. After washing and storage at 4 degrees C for 24 hour, the red cells had a mean freeze-thaw-wash recovery value of 90 percent, a mean 24-hour posttransfusion survival value of 85 percent, a mean index of therapeutic effectiveness of 75 percent, normal or slightly impaired oxygen transport function, and minimal hemolysis.(ABSTRACT TRUNCATED AT 250 WORDS)

The bottom and top system: a new technique for blood component preparation and storage.

A new, automated technique for the preparation of blood components is described. A system of 3 or 4 integrally connected plastic containers (Optipac) is handled by a new type of extractor (Optipress). The container in which the blood is collected has an outlet at the top and another at the bottom. After normal centrifugation to obtain separation of the blood components, these are squeezed out from the top and bottom simultaneously under control of a photocell. The primary separation step results in three components: a leukocyte-poor red-cell suspension in SAGM medium, CPD plasma, and a buffy-coat preparation. The system has been tested in two laboratories (lab A and lab B). A 'heavy-spin' centrifugation to obtain a maximum yield of cell-poor plasma gave the best removal of leukocytes from the red cells; the remaining leukocyte content was 0.46 +/- 0.25 (lab A) and 0.5 +/- 0.4 (lab B) x 10(9)/red-cell unit. Platelet concentrates can be prepared either the normal way via platelet-rich plasma or from buffy coat. Red-cell 24-hour autologous posttransfusion survival using labeling with 51Cr was 87.5 +/- 4.1% (lab A) after 35 days, and 84.2 +/- 4.2% (lab A) and 77.5 +/- 1.5% (lab B) after 42 days. Red-cell morphology and fluidity compared favorably to previous studies using the same additive solution in traditional plastic-bag systems. The total adenine nucleotide concentration was maintained normal for 42 days.(ABSTRACT TRUNCATED AT 250 WORDS)

The 24-hour posttransfusion survival and lifespan of autologous baboon red cells treated with inositol hexaphosphate-polyethylene glycol or inositol hexaphosphate-adenosine triphosphate-polyethylene glycol to decrease oxygen affinity.

Baboon red cells were treated to reduce oxygen affinity by an osmotic-pulse procedure using dimethyl-sulfoxide. Inositol hexaphosphate (IHP) alone or IHP and adenosine triphosphate (ATP) were incorporated into the red cells in the presence of polyethylene glycol (PEG). The procedure produced variable increases in the red cell P50 value, i.e., the partial pressure of oxygen at which 50% of the hemoglobin was saturated. The effect of treatment of autologous baboon red cells on the 24-hour posttransfusion survival value and lifespan T50 value was measured using a double-label procedure. The data demonstrate that the increase in the P50 value of treated red cells was negatively correlated with the 24-hour posttransfusion survival value; the higher the P50 value, the poorer the 24-hour posttransfusion survival value. The 24-hour posttransfusion survival value for nontreated baboon red cells was 90% and the T50 value was 14 days. The IHP-ATP-PEG-treated red cells had significantly higher red cell ATP levels than did IHP-PEG-treated red cells. The 24-hour posttransfusion survival value was 68% for the IHP-ATP-PEG treated red cells and 52% for the IHP-PEG-treated red cells when the increase in P50 ranged from 10 to 20 mm Hg; the lifespan T50 value for both the IHP-ATP-PEG-treated red cells and the IHP-PEG-treated red cells was 15 days. Osmotic pulse treatment produced significant red cell injury manifested by the 24-hour posttransfusion survival value. However, modification of the RBC with IHP-PEG or IHP-ATP-PEG to decrease hemoglobin affinity for oxygen did not affect their lifespan.

アスコルビン酸２‐リン酸（ＡＰ）を含む赤血球保存液

Plasma and leukocyte poor red cell concentrates were stored in a new additive solution containing mannitol, citrate, phosphate, adenine and ascorbyl-2-phosphate for 6 weeks. The additive solution, MAAP-2 showed the best results.The 24-hour autologus post-transfusion survival of 51Cr-labeled red cell was 95.1% and 89.3% for 3 and 6 weeks storage, respectively.After 6-week stroage, ATP and 2, 3-DPG levels were 2.96μmole/gHb and 7.47μmol/gHb, respectively. Futhermore, the stored red cell shapes were ratained discoid and hemolysis decreased.This solution enabled to provide an excellent red cell product with a long shelf life and with a normal oxygen affinity.

The 24-Hour Posttransfusion Survival and Lifespan of Autologous Baboon Red Cells Treated with Inositol Hexaphosphate-Polyethylene Glycol or Inositol Hexaphosphate-Adenosine Triphosphate-Polyethylene Glycol to Decrease Oxygen Affinity

Baboon red cells were treated to reduce oxygen affinity by an osmotic-pulse procedure using dimethylsulfoxide. Inositol hexaphosphate (IHP) alone or IHP and adenosine triphosphate (ATP) were incorporated into the red cells in the presence of polyethylene glycol (PEG). The procedure produced variable increases in the red cell P(50) value, i.e., the partial pressure of oxygen at which 50% of the hemoglobin was saturated. The effect of treatment of autologous baboon red cells on the 24-hour posttransfusion survival value and lifespan T(50) value was measured using a double-label procedure. The data demonstrate that the increase in the P(50) value of treated red cells was negativelycorrelated with the 24-hour posttransfusion survival value; the higher the P(50) value, the poorer the 24-hour posttransfusion survival value. The 24-hour posttransfusion survival value for nontreated baboon red cells was 90% and the T(50) value was 14 days. The IHP-ATP-PEG-treated red cells had significantly higher red cell ATP levels than did IHP-PEG-treated red cells. The 24-hour posttransfusion survival value was 68% for the IHP-ATP-PEG treated red cells and 52% for the IHP-PEG-treated red cells when the increase in P(50) ranged from 10 to 20 mm Hg; the lifespan T(50) value for both the IHP-ATP-PEG-treated red cells and the IHP-PEG-treated red cells was 15 days. Osmotic pulse treatment produced significant red cell injury manifested by the 24-hour posttransfusion survival value. However, modification of the RBC with IHP-PEG or IHP-ATP-PEG to decrease hemoglobin affinity for oxygen did not affect their lifespan.

Studies on the mechanism of human red cell loss of viability during storage at +4 degrees C in vitro. I. Cell shape and total adenylate concentration as determinant factors for posttransfusion survival.

Red cells stored in SAGM medium for 42 days at +4 degrees C were rejuvenated by bicarbonate, pyruvate and adenosine. Autologous 24-hour posttransfusion survival was determined in untreated as well as rejuvenated cells and showed an improvement from 77.4 +/- 4.7 to 89.2 +/- 7.2%. The erythrocyte adenylate energy charge decreased relatively more than the total adenylate concentration during storage, but the latter correlated better with posttransfusion red cell survival. Considerable deteriorations in red cell morphology (expressed as morphology index) and in deformability (measured as red cell fluidity) were observed during storage but were partly reversed by rejuvenation. The morphology index and the posttransfusion survival showed a significant correlation (r = 0.95, p less than 0.005) after, but not before, rejuvenation, indicating that the remaining changes are more permanent and decisive of survival. It is suggested that, in the proportion of stored erythrocytes which respond to rejuvenation, the capacity and time dependence of recovery of normal shape and flexibility are important.