Single-donor Platelet Concentrates Research Articles

Predicting Platelet Age Using Artificial Intelligence Techniques

Background: Platelets are small, enucleated cells with their primary physiological role to repair vascular damage (hemostasis) and initiate thrombus formation in response to vascular injury. Low platelet count (thrombocytopenia) is associated with increased bleeding risk and is caused by decreased platelet production and/or increased platelet clearance. Platelet transfusion is a routinely used lifesaving procedure to control or prevent bleeding in patients with thrombocytopenia or platelet dysfunction. During storage platelet function deteriorates significantly over time and regulation of platelet granular content is essential in maintaining a healthy platelet function. Currently no in vitro tests are available to predict platelet age, which may be used to diagnose the underlying cause of thrombocytopenia, or to determine storage time of platelet concentrates (PC). Using confocal and super-resolution microscopy we could quantify loss of platelet granule content during aging. In this study we used Artificial intelligence techniques (AIT) to derive exact platelet age from imaging data. Aim: To develop AIT-based algorithms that can determine platelet age from images of platelets stored in plasma-rich-plasma (PRP) and in platelets concentrates (PC) measured by confocal microscopy. Methods: Platelets were stored in PRP for 8 hours at room temperature. Platelets were isolated and stained with antibodies for alpha-tubulin, von Willebrand Factor (VWF), SPARC or alpha-tubulin and imaged at T=0, 4 and 8 hours using confocal microscopy. Additionally 3 single donor PC in plasma were stored for 10 days at standard blood bank conditions. Platelet samples were pooled and analyzed at day 1,2,3,6,7,8,9 and 10. We trained a neural network consisting of multiple convolutional layers to classify age (time after donation) of platelets that were stored in either PRP or PC. Results: During storage in PRP as well as of PC we used for each time point 1334 62µm x 62µm x 4µm 3D images, containing 50 to 150 platelets per image, for training. When staining with markers VWF, SPARC or alpha-tubulin alone as training set we could already classify mean platelet age up to 78%, 85%, 88% respectively. When combining all three markers a maximum accuracy of 95% for platelets stored in PRP up to 8 hours was reached (Figure 1). The additional effect of combining these three markers highlights that the model used information in the images synergistically. For PC, an accuracy of 90% was reached for classifying mean platelet age of platelets stored up to 8 days. The model could not be trained to discriminate between days 9 and 10 resulting in an overfitted model. However the model was able to distinguish day between day 1, 3, 6 and 10 (Figure 2). As negative control a model trained on a randomized sample was made. This control model was not able to assign a category correctly, indicating an actual difference in platelet characteristics during storage was detected. By using visualization of the weight-gradient (e.g. where in the images the model assigns the most weight) we additionally confirmed that the model used separate individual platelets to characterize the age of the complete platelet sample. Conclusion: Combining confocal platelet images with a convolutional neural network, we trained a model that could synergistically integrate different stainings and information of individual platelets to predict with >95% accuracy platelet age in vitro as well as in PC up to 10 days of storage. Additionally our model could distinguish "young" platelets (stored 1, 3 or 6 days) from "old" platelets (stored 10 days). Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

A prospective observational study to compare transfusion outcomes in ABO identical versus ABO non-identical single donor platelet concentrates: An experience from a tertiary healthcare center in India.

ABO incompatible single donor platelet concentrates (SDPC) have a concern about unsatisfactory increments as well as possibility of hemolytic transfusion reaction. But from Indian population no study has commented on the clinical and laboratory outcome of ABO mismatched platelet transfusion. The aim of study was to compare transfusion outcomes in ABO identical versus ABO non-identical single donor platelet concentrates. In this prospective observational study, 400 SDPC transfusions among different patients were included. In group A (n=200), ABO identical SDPC transfusions and in group B (n=200) ABO non-identical SDPC transfusions were added. Corrective count increment (CCI), absolute count increment (ACI), percent platelet recovery (PPR) were calculated and incidents of hemolytic transfusion reactions were noted. In group A mean±SD of ACI, CCI and PPR were as 30.78±12.51, 15.10±6.677, 39,948.9±20,099.392. In group B, mean±SD of ACI, CCI and PPR were-25.4±15.65, 12.509±5.906, 33,559.2±22,150.304. And when CCI, ACI, PPR were compared with group A and group B, statistically significant differences were noted (P<0.05). There was statistically significant difference in CCI, ACI and PPR in oncology patients and other prophylactic recipients except patients with dengue and other infectious disease. But there was no hemolytic transfusion reaction noted in any group. Our study clearly establish the potential benefits of ABO-identical PLT transfusion. It also points out that in emergency conditions or when there is a paucity in inventory, ABO non-identical SDPC transfusion may be lifesaving and clinically significant.

Frequency and clinical characteristics of allergic transfusion reactions in children

Allergic transfusion reactions (ATRs)are a common form of acute transfusion reaction. It was aimed to determine the clinical characteristics and frequency of ATRs in children. This study included children who were transfused with red cell concentrate (RCC), fresh-frozen plasma (FFP), platelet concentrates(PC), apheresis granulocyte, and cryoprecipitate.The patients’ sociodemographic characteristics, the blood product that caused the reaction, the type and timing of the reaction, the patient’s age at time of reaction and their diagnosis, follow-up period, and clinical data were recorded. A total of 89703 bags of blood products were transfused to 4193 children.Two hundred eleven acute transfusion-related reactions occurred in 157 (3.74%) patients.Of these, 125 reactions (59%) were allergic. ATR occurred in 125 of 89703 infusions (0.14%).The median age of patients was 9.99 years (IQR:4.67-14.38) and ATRs occurred at a median of 30 minutes into the transfusion. Eighteen (18%) of the patients also had a history of drug reaction.When the blood products that caused ATRs were examined, 43(34.5%) occurred with apheresis and single-donor PC, 37(29.6%) with FFP, 32 (25.6%) with RCC, 10(8%) with pooled PC, 2(1.6%) with cryoprecipitate, 1(0.8%) with apheresis granulocyte.Ninety-nine(79%) of the reactions were minor allergic reactions and 26(21%) were anaphylaxis.Compared to minor allergic reactions, the proportion of PCs was statistically higher in anaphylaxis(p=0.02). Patients receiving PC should be monitored more carefully during the first half hour of transfusion. In addition, approximately one-fifth of the patients who developed ATR also had a history of drug reaction. Patients with previous reactions to drugs may be more likely to have ATR.

Determination of thromboelastographic responsiveness in stored single‐donor platelet concentrates

Thromboelastography (TEG) is widely used in hospitals but less commonly in blood banks for evaluation of platelet (PLT) concentrates (PCs). A TEG-PC assay for testing fresh or stored PLTs must reflect the quality of the PLTs. The added value could be measurement of donor-dependent PLT quality. Whole blood (WB) normal values were generated from 100 donors, using standard tests. Nineteen single-donor PCs were evaluated with a TEG-PC assay, using Octaplas as microparticle-free diluent and kaolin or collagen as activator, stored up to 12 days, and also sampled for additional in vitro tests. WB values showed larger reaction rates (R- and K-times, angle) compared to the reference values and almost similar maximum amplitude (MA). PCs showed usual storage lesion and TEG-PC results showed significant decreasing R- and K-times and increasing angle. Mean MA values remained constant but individual measurements were affected by clot retraction. TEG tracings of two PCs with good quality on Day 12 showed weak to strong clot retraction, while two PCs with poor quality showed moderate clot retraction on Day 1, no clot retraction on Days 5 to 12, and a decreased MA on Day 12. Clot strength (MA) and especially clot retraction represent possibly donor-specific effects. A TEG-PC assay has been developed that is sensitive to storage effects. The assay has the potential to be helpful in selection of PLT donors but needs improvement to be more sensitive, reproducible, and distinctive to determine whose PLTs store poorly and whose store well.

Evaluation of Different Preparation Procedures of Pathogen Reduction Technology(Mirasol®)-Treated Platelets Collected by Plateletpheresis

Background: The Mirasol® pathogen reduction technology (PRT) for platelet concentrates (PC) uses riboflavin and UV light (270–360 nm). We evaluated the impact of PRT on platelets in comparison to standard single-donor PC. Material and Methods: Platelets were resuspended in autologous plasma. After 2 h rest without agitation, PC were split into an untreated control unit (C-PC) and an immediately treated unit (T-PC) (series I). In series IV, split PC were stored under agitation over night before PRT was carried out. Platelet quality was assessed by pH, glucose consumption, lactate production rate, LDH, soluble sCD62p and CD62p expression with and without TRAP (thrombin receptor-activating peptide) over 7 days. Results: Series I: On day 5, pH values were lower for T-PC (6.8 ± 0.2 vs. 7.4 ± 0.1, C-PC), accompanied by a higher glucose consumption rate of 0.069 ± 0.016 vs. 0.035 ± 0.006 mmol/10¹² platelets/h and lactate production rate of 0.126 ± 0.031 vs. 0.063 ± 0.011 mmol/10¹² platelets/h. CD62p using TRAP was lower for T-PC (50 ± 11 vs. 62 ± 14%). Baseline activation was higher in T-PC (35 ± 12 vs. 28 ± 15%). Longer initial rest time had no impact on these results (series II/III/IV). Conclusion: PRT leads to an increase of platelet metabolism and activation independent of the length of the initial rest times. PC resuspended in autologous plasma should be stored at maximum up to day 5.

In vitro evaluation of metabolic changes and residual platelet responsiveness in photochemical treated and gamma‐irradiated single‐donor platelet concentrates during long‐term storage

Photochemical treatment (PCT) prevents replication of pathogens in platelet concentrates (PCs) by cross-linking nucleic acids and thus affects all cells containing DNA or RNA. Fourteen double-dose single-donor PCs were divided into two study arms. The double-dose PCs were split in two identical units, PCT and conventional control PCs. Study Arm A consisted of seven PCT PCs with corresponding untreated controls, whereas Study Arm B consisted of seven PCT PCs with corresponding gamma-irradiated control. Metabolic changes and agonist-induced platelet (PLT) response were evaluated during storage for up to 12 days. Higher rate of PLT destruction, illustrated by reduced PLT content, increased lactate dehydrogenase levels, and higher CD61+ microparticle formation rate, were observed after PCT. Generally PCT accelerated metabolic changes in PCs and reduced agonist-induced (collagen or thrombin receptor activator peptide [TRAP]) aggregation responses. Flow cytometric analysis of CD62P and CD42b (GPIbalpha) expression showed higher spontaneous PLT activation in PCT PCs from 5 days of storage. Correspondingly, a reduced capacity for up regulation of CD62P expression and down regulation of CD42b was observed in PCT PLTs after stimulation by the agonists ADP or TRAP. Generally reduced in vitro PLT quality was observed after PCT during storage for up to 12 days, with marked reduction from 5 days of storage. Compared to conventional PCs, reduced agonist-induced aggregation and glycoprotein expression were observed after PCT during storage, corresponding to significantly higher level of spontaneous PLT activation in PCT PCs. Clinical studies of efficacy and safety of PCT PCs stored for more than 5 days are recommended.

Bacterial contamination of platelet concentrates prepared by different methods: results of standardized sterility testing in Germany

National guidelines for monitoring the bacterial contamination rate of blood components were introduced in Germany in 1997. The objective of this study was to present and evaluate the results of sterility testing of platelet concentrates (PCs) prepared by different methods. The analysis of results of sterility testing of blood component production from transfusion medicine centres in Germany in 1998 and 2001 was based on information collected using standardized questionnaires. The bacterial contamination rates for single-donor PCs derived from whole blood and apheresis (0.210% vs. 0.156%) were comparable and showed no significant difference. However, pooled PCs produced from four buffy coats using the sterile docking procedure showed a significantly higher bacterial contamination rate compared with single-donor PCs derived from whole blood and apheresis (0.184% vs. 0.604%). Use of standardized methods for sterility monitoring is sufficient to assess collection and production processes in terms of hygiene and yields reliable data on bacterial contamination rates of blood components. The methods described are suitable for using to analyse the efficiency of newly introduced methods to reduce bacterial contamination rates of blood components (e.g. diversion, bacteria screening and pathogen inactivation).

Fresh frozen plasma and platelet concentrates may increase plasma anti-diphtheria toxin IgG concentrations: Implications for diphtheria fusion protein therapy.

We are developing two fusion proteins consisting of a diphtheria toxin (DT) linked to either granulocyte macrophage colony stimulating factor (DT388-GMCSF) or interleukin-3 (DT388-IL3). In trials, patients with anti-DT IgG concentrations >2.5 microg/ml had significantly lower concentrations of either fusion protein. DT389-IL2 is currently FDA approved for the treatment of cutaneous T-cell lymphomas. We noted increased concentrations of anti-DT IgG after administration of platelet concentrates (PC). Because many of these patients require transfusions, we measured the anti-DT IgG content of FFP and PC. We assayed 14 bags of FFP and 12 bags of single-donor PCs for anti-DT IgG by an enzymoimmunoassay against DT389-IL2, DT388-IL3, DT388-GMCSF. The median (percent of samples positive) of anti-DT IgG concentrations in PC against DT388-GMCSF, DT388-IL3, DT389-IL2 was 0.8 microg/ml (83%), 0.7 microg/ml (83%), and 0.3 microg/ml (58%), respectively. The median (percent of samples positive) anti-DT IgG concentration in FFP against DT388-GMCSF, DT388-IL3, and DT389-IL2 was 2.1 microg/ml (86%), 1.9 microg/ml (93%), and 1.4 microg/ml (86%), respectively. There was a strong association between anti-DT389IL2 IgG, anti-DT388IL3 IgG, and anti-DT388GMCSF IgG concentrations in both the FFP (95.6%) and PC (76.3%). Assuming a plasma volume of 3 l in a 70 kg patient, a single FFP unit would increase the plasma anti-DT389IL2 IgG, anti-DT388IL3 IgG, and anti-DT388GMCSF IgG by 0.13 microg/ml, 0.17 microg/ml, and 0.19 microg/ml, respectively. For PC, a single unit would increase plasma anti-DT389-IL2 IgG, anti-DT388-IL3 IgG, and anti-DT388-GMCSF IgG by 0.03 microg/ml, 0.06 microg/ml, and 0.07 microg/ml, respectively. In conclusion, a single FFP or PC appears to minimally increase anti-DT IgG concentrations, but multiple units may significantly do such.

Leukocyte Depletion and Storage of Single-Donor Platelet Concentrates

Background and objectives: Because of widespread use of leukocyte reduction in platelet concentrates (PCs) and the need to store such concentrates, we investigated the effects of leukocyte depletion on the quality of stored PCs. Materials and methods: Ten double-sized PCs were divided into 2 equal units which were tested simultaneously. One half was stored for 5 days after filtration through a polyester filter, the other one was stored unfiltered. Results: The volume of the 10 ‘oversized’ PCs was 483±40ml (mean ± standard deviation) and they contained 5.9± 1.5×10^11 platelets and 80±23×10^6 leukocytes. Filtration significantly reduced the leukocyte concentration (168±56/pl before, 6±4/μl after filtration) and leukocyte count (39.9±11.3×10^6 vs. 1.3±0.9×10^6; p<0.0005). Filtration caused a platelet loss of 16%, the platelet count decreasing not significantly from 2.91±0.75×10^11 to 2.40±0.94×10^11 (p = 0.26). After 5 days of storage all parameters of platelet function (platelet aggregation to several stimuli, hypotonic shock reaction [HSR] and platelet retraction), mean platelet volume, and pH and pCO(2) showed no advantage for PCs filtered prior to storage compared to PCs stored unfiltered. Moreover, platelet aggregation on day 5 using 4 agonists at 10 concentrations showed worse results in 4 assays in prestorage filtered PCs (collagen [4μg/ml: p<0.05, ADP [0.2 mM]: p<0.05, ADP [0.3 mM\\ p<0.05, thrombin [0.6 E/ml] : p<0.05). But there is no convincing trend in all aggregation tests, and HSR, presumably the most useful parameter, was not different on day 5. Conclusions: There is no advantage in terms of improved quality for prestorage leukodepletion of PCs. Taking into account the obvious disadvantages of filtration, such as platelet loss and increasing costs per transfusion, we conclude that pre- or post-storage filtration of single-donor PCs should be done only for patients who have a clear indication for the transfusion of leukocyte-poor blood products.

Leukocyte depletion and storage of single-donor platelet concentrates.

Because of widespread use of leukocyte reduction in platelet concentrates (PCs) and the need to store such concentrates, we investigated the effects of leukocyte depletion on the quality of stored PCs. Ten double-sized PCs were divided into 2 equal units which were tested simultaneously. One half was stored for 5 days after filtration through a polyester filter, the other one was stored unfiltered. The volume of the 10 "oversized' PCs was 483 +/- 40 ml (mean +/- standard deviation) and they contained 5.9 +/- 1.5 x 10(11) platelets and 80 +/- 23 x 10(6) leukocytes. Filtration significantly reduced the leukocyte concentration (168 +/- 56/microliter before, 6 +/- 4 /microliter after filtration) and leukocyte count (39.9 +/- 11.3 x 10(6) vs. 1.3 +/- 0.9 x 10(6); p < 0.0005). Filtration caused a platelet loss of 16%, the platelet count decreasing not significantly from 2.91 +/- 0.75 x 10(11) to 2.40 +/- 0.94 x 10(11) (p = 0.26). After 5 days of storage all parameters of platelet function (platelet aggregation to several stimuli, hypotonic shock reaction [HSR] and platelet retraction), mean platelet volume, and pH and pCO2 showed no advantage for PCs filtered prior to storage compared to PCs stored unfiltered. Moreover, platelet aggregation on day 5 using 4 agonists at 10 concentrations showed worse results in 4 assays in prestorage filtered PCs (collagen [4 micrograms/ml: p < 0.05, ADP [0.2 mM]: p < 0.05, ADP [0.3 mM]: p < 0.05, thrombin [0.6 E/ml]: p < 0.05). But there is no convincing trend in all aggregation tests, and HSR, presumably the most useful parameter, was not different or day 5. There is no advantage in terms of improved quality for prestorage leuko-depletion of PCs. Taking into account the obvious disadvantages of filtration, such as platelet loss and increasing costs per transfusion, we conclude that pre- or post-storage filtration of single-donor PCs should be done only for patients who have a clear indication for the transfusion of leukocyte-poor blood products.

Economic Impact of Using Single-Donor Platelets

Rapid growth of platelet production in Bristol, UK, and the requirement for a maximum 8-hour hold time prior to the production of platelets from whole blood led to the use of single donor platelet concentrates (SDPC). Initially platelets were derived from platelet rich plasma (PRP) produced by the Haemonetics system. The COBE Spectra system was introduced based on the high yield and low leucocyte content of the Spectra product and the absence of required secondary processing. Platelets could be produced more cheaply on the Haemonetics system if plasma revenues were included, however, when the premium paid for apheresis plasma was eliminated and when secondary processing costs were taken into account, the COBE platelets became less expensive. Buffy-coat platelets from whole blood, because of their higher yield and reduced leucocyte levels, have replaced platelets derived by the PRP method. Based on a costing model which assigns all collection and testing costs to red blood cells (RBCs), the key product or cost driver, buffy-coat platelets now appear to be very inexpensive in comparison to SDPC. Because the current costing models do not take into account patient outcome, further research into this area is needed.

Single-donor Platelet Concentrates Stored in Synthetic Medium. In Vitro and in Vivo Studies

Single-donor platelet concentrates (PC) were prepared in 80-120 ml plasma and stored in two polyolefin bags after addition of 250 ml plasmalyte, a simple, glucose-free synthetic medium that was previously used for platelet storage; when compared to PC stored in plasma, PC stored in plasmalyte, showed similar platelet quality, morphology and function after 5 days of storage. In vivo increments observed after transfusion of PC stored for 5 days in plasmalyte were similar to those observed after transfusion of 1-2 day old PC stored in plasma. Moreover, transfusion of 5-day old PC stored in plasmalyte was associated with correction of prolonged bleeding times in all 3 of the 3 patients evaluated. It is concluded that plasmalyte seems to be promising as a medium for single-donor PC storage.

Morphological changes in thrombocytes during blood bank storage. An ultrastructural morphometric study

Monitoring the storage lesion of platelet concentrates (PC) is commonly done by determining metabolic parameters, release of cytoplasmic markers and granules contents and watching the light microscopical appearance of platelets. Descriptions of ultrastructural changes are rare, and so we ascertained the changes in two important organelles, the alpha- and lysosomal granules and the open canalicular system (OCS), by morphometric analysis. Single donor PC's were prepared with a CS-3000 plus (Baxter) processor and stored under routine conditions. The volume fraction of the granules displayed a decrease to 43% after 8 days of storage, and the OCS fraction increased to 143%. Moreover, changes in the morphology of granules and the development of OCS channels are described, and ballooning and even complete lysis of some platelets could be observed after only three days. Furthermore, the membrane flux in the formation of pseudopodia is discussed. Our results stress the need to establish better preparation and storage conditions for platelet concentrates.

Storage of single‐donor platelet concentrates: metabolic and functional changes

During the last decade, the trend toward intensifying chemotherapeutic regimens in patients with hematologic malignancies rapidly increased the demand for single-donor platelet concentrates (PCs). The logistics of such supply, however, necessitated the storage of these blood components prior to transfusion. Today, most blood centers use di(2-ethylhexyl)phthalate-free blood bags, which are assumed to allow a storage period of up to 5 days. This report describes biochemical and functional changes of stored single-donor PCs, which may influence the expected quality of PCs. The acid-base status is characterized by an initial respiratory alkalosis compensated by a metabolic acidosis. Changes in extracellular electrolyte, lactate dehydrogenase, glucose, lactate, elastase, and complement levels, as well as in the release of alpha granule content and the initial activation of plasma coagulation, are demonstrated. These changes result in a functional impairment of stored PCs as reflected by thromboxane and serotonin release reaction and by aggregation and in vitro bleeding time studies. In contrast, in vivo recovery and survival rates have been reported to be unaffected. Whether the good recovery and survival rates are caused by a rejuvenescence of stored PCs in vivo or are due to injured circulating platelets has not yet been proven.