A Comparative Study in Volunteers of Apheresis and Buffy Coat Derived Platelets

It is important that, at the time of transfusion, platelets prepared by different techniques are effective in vivo and meet acceptable criteria. A paired study was undertaken in 8 volunteer donors to compare apheresis platelets collected on the COBE Spectra with platelets derived from the buffy coat of whole blood donations after 5 days storage. In vivo recovery, survival and biodistribution were determined following indium-111 labelling of the platelets and autologous infusion into volunteers together with the in vitro evaluation of platelet function and biochemistry.

The in vivo and in vitro characteristics of both types of platelet concentrate were not significantly different. Both products were equally viable after 5 d storage and both were of an acceptable quality as judged by currently used platelet products. Mean platelet survival (multiple hit) was 5.4 d for the apheresis platelets and 6.9 d for the buffy coat derived platelets and maximum recovery in circulation was 28.1% and 34.8%, respectively. There was a significantly higher platelet yield, as expected, from apheresis and a 1.5 log lower level of white cell contamination. This would be advantageous for patients in need of prolonged or recurrent transfusions as the number of donor exposures and the risk of alloimmunisation or viral transmission would be reduced.     

Survival in vivo of platelets stored for 48 hours in the buffycoat at 4°C compared to platelet rich plasma stored at 22°C

Summary High speed centrifugation allows separation of whole blood into cell free plasma, a buffy coat and leukocyte poor red cells. The buffy coat can be used for the preparation of platelet concentrates. High lactate production at 22°C requires storage of the buffy coat at 4°C. Survival in vivo of platelet concentrates prepared from buffy coats stored at 4°C for 48 h (BC-PC) was compared with the survival in vivo of platelet concentrates from platelet rich plasma stored at 22°C for 48 h (PRP-PC). Both methods were studied in the same healthy volunteers (n=8) using⁵¹Cr labeled autologous platelets. The mean ±SD recovery 15 min after reinfusion of the BC-PC was 30.5%±13.3% and for PRP-PC 41.4%±7.9% (p<0.0001). The survival in vivo for BC-PC was 2.4 days ±0.4 days and for PRP-PC 7.0 days ±1.4 days (p<0.0001).Since the survival in vivo is significantly less for platelets derived from the buffy coat stored at 4°C, we advocate storage of platelets at 22°C.     

High platelet content can increase storage lesion rates following Intercept pathogen inactivation primarily in platelet concentrates prepared by apheresis

Background

Pathogen inactivation methods for platelet concentrates are increasingly being used in blood banks worldwide. In vitro studies have demonstrated its effects on storage lesion, but little routine quality control data on blood banking outcomes have been reported.

Materials and Methods

Swirling of distributed products was monitored before and after implementation of Intercept pathogen inactivation. Metabolic parameters pH, glucose and lactic acid were determined in a random cohort of expired pathogen‐inactivated products. Storage lesion indicators in apheresis concentrates with premature low swirling were compared to concentrates with normal swirling.

Results

During validation for implementing Intercept pathogen inactivation, pH and glucose levels decreased faster in apheresis platelet concentrates with high platelet content than with low platelet content or than in pathogen‐inactivated pooled buffy coat‐derived products. In routine products, glucose exhaustion was more often found in apheresis compared to buffy coat‐derived platelet concentrates despite 3–7% more plasma carryover in the former. Annual incidence of premature low swirling increased significantly by 50% following implementation of pathogen inactivation implementation for apheresis but not for pooled buffy coat platelet concentrates. In addition, apheresis concentrates with premature low swirling had a significantly higher median platelet count (5·0 × 10¹¹) than unaffected products (3·5 × 10¹¹).

Conclusion

The risk of increased storage lesion rates following Intercept pathogen inactivation is higher for apheresis than for buffy coat‐derived platelet concentrates, especially when platelet contents are higher than 5·0 × 10¹¹.     

A critical comparison of platelet preparation methods

PURPOSE OF REVIEW: Platelet concentrates may be prepared from whole blood or by plateletpheresis. Currently, the non-evidence-based preponderance of apheresis units in the United States and the 50: 50 ratio in Europe may not optimize the gifts of whole-blood donors or minimize healthcare costs. Post-storage pooled, whole-blood-derived platelets, on the other hand, do not provide the convenience of or an equivalent level of safety as apheresis platelets. RECENT FINDINGS: Some data suggest that different methods of manufacture of whole-blood-derived platelets (platelet-rich plasma or buffy coat intermediate steps) result in differing degrees of platelet activation, which may impact on the quality of stored concentrates. Recent studies have observed superior radiolabel recovery and post-transfusion increments for platelets derived from apheresis compared with platelet-rich plasma whole-blood-derived platelets. A pre-storage pooling system for whole-blood-derived platelets has just been licensed in the USA, and may eventually combine the benefits of apheresis-derived and whole-blood-derived platelets. The advantages of the European method of manufacture of buffy coat whole-blood-derived platelet concentrate have convinced the Canadian Blood Services to abandon platelet-rich-plasma-derived concentrates. SUMMARY: We present a literature-based review of the relative merits of apheresis-derived and whole-blood-derived platelets. Additional studies are needed in order to define the optimal proportion of the platelet supply from apheresis collections and the choice of whole-blood-derived production method for US blood providers.     

The Mirasol? Pathogen Reduction Technology system and quality of platelets stored in platelet additive solution

Background

The Mirasol Pathogen Reduction Technology system for platelets and plasma uses riboflavin and UV light to introduce irreparable lesions into nucleic acids thereby inhibiting pathogen and white blood cell replication and reducing the load of infectious pathogens. The aim of the present study was to evaluate low plasma buffy coat platelet concentrates obtained from the OrbiSac System and to examine the effects on the development of platelet storage lesion during storage in platelet additive solution.

Material and Methods

Twenty buffy coat platelet concentrates were generated by pooling five individual units using the OrbiSac System. Riboflavin was added during the final pooling step, and the units were exposed to UV light. The bag was removed after the target energy of 6.24 J/mL had been delivered and 150 mL of platelet additive solution were added prior to storage. Platelet quality was assessed by pH, swirl, CD62P expression, lactate dehydrogenase, lactate production and glucose consumption rates over 7 days of storage.

Results

Buffy coat platelet concentrates generated on the OrbiSac contained an average 3.5 ± 0.6 x 10¹¹ platelets at a concentration of 2976± 406 x 10⁶/mL. After addition of 150 mL platelet additive solution the storage concentration was 1043 ± 148x 10⁶/mL. Values obtained for pH, lactate production and glucose consumption rates were all within the limits of previously established correlations between in vitro cell quality and in vivo performance of Pathogen Reduction Technology-treated platelets in plasma.

Discussion

In vitro studies show that OrbiSac-derived platelets treated with the Mirasol Pathogen Reduction Technology system preserve adequate function, which would indicate acceptable in vitro viability.     

Platelets Stored in a Glucose-Free Additive Solution or in Autologous Plasma – An Ultrastructural and Morphometric Evaluation

We evaluated a new glucose-free citrate-acetate-NaCl platelet additive solution (PAS 2). In series I, platelet concentrates (PC) were prepared by apheresis and subsequently stored either in plasma (n =16) or in PAS 2 (n =15). In series II, PCs were prepared from pools of four buffy coats (BC) and stored in plasma (n =12) or in PAS 2 (n =11). By means of ultrastructural morphometry, the volume fractions of α-granules, the open canalicular system (OCS) and the fraction of storage granules secreted into the OCS were analyzed during storage for up to 8 days. Additionally, we determined pH, glucose, lactate, pCO₂, HCO₃^–, lactate dehydrogenase and platelet factor 4. Apheresis platelets stored in plasma showed no changes in their contents of α-granules and in the fractions of the OCS. In contrast, apheresis platelets stored in PAS 2 displayed a decrease of their relative volume fraction of α-granules from 9.1±1% on day 1 to 3.7±0.9% on day 5. The fraction of the OCS increased from 7.4±0.8% on day 1 to 17.1±1.4% on day 3. On day 8, 93±9% of all platelets were lysed. Levels of glucose were significantly lower in these preparations and after day 3 glucose consumption decreased to zero. Among PC derived from pooled BC, differences between storage in PAS 2 or plasma were less striking. Only the fraction of α-granules secreted into the OCS was significantly greater in BC derived PC stored in PAS 2 on all days. These PCs stored in PAS 2 had a higher plasma carryover (30%) in comparison to apheresis PC stored in PAS 2 (10%). We conclude that plasma is superior to PAS 2 for storage of both apheresis and buffy coat platelets. For preservation of the structural integrity of platelets, the use of PAS 2 requires a minimum of 30% plasma carryover.     

Storage of platelets in additive solutions: a pilot in vitro study of the effects of potassium and magnesium

BACKGROUND AND OBJECTIVES: Platelet additive solutions (PAS) have been shown to be suitable for extended platelet storage but have required the carryover of substantial (30%) amounts of plasma for success. Improving platelet quality by optimizing the composition of PAS may allow a reduction to be made in the amount of plasma carried over. Reducing the proportion of plasma carried over would facilitate some methods of viral inactivation and make available greater amounts of plasma for other needs. MATERIALS AND METHODS: Platelets from six pools of 25 buffy coat platelet units and five apheresis platelet units were aliquoted for storage in plasma, or converted to PAS units in either a specific additive solution (PAS-III), with 30% or 20% plasma, or a modification of PAS-III containing 5.0 mm potassium and 1.5 mm magnesium (PAS-IIIM), with 30% or 20% plasma. Units were stored at room temperature with agitation for 7 days with in vitro testing for biochemical, haematological and functional parameters. RESULTS: Storage of platelets in PAS-IIIM resulted in a reduced rate of glycolysis and better retention of pH, morphology score and ATP levels. Platelets initially showed less evidence of activation when stored in PAS-IIIM, with reduced P-selectin expression. Storage in PAS-IIIM with 20% (rather than the standard 30%) plasma appeared to result in the retention of in vitro properties, similarly to storage in standard PAS-III with 30% plasma. CONCLUSIONS: Storing platelets in an additive solution containing magnesium and potassium improves the functionality of the platelets, as measured by in vitro testing, and may allow a reduction of the amount of plasma required to be carried over to the final unit.     

Cytokine levels as performance indicators for white blood cell reduction of platelet concentrates

BACKGROUND AND OBJECTIVES: With the implementation of universal white blood cell (WBC) reduction in the UK, in-process WBC-reduction filters for pooled buffy coat (BC)-derived platelet concentrates (PCs) and apheresis methods are used routinely for the production of WBC-reduced PCs. While these strategies meet the specification for WBC reduction (< 5 x 10(6) WBCs/unit), the products from these processes may differ depending on the process employed and its performance. The aim of this study was therefore to investigate whether PCs prepared using various WBC-reduction processes are sufficiently depleted of WBCs to limit cytokine accumulation during storage and to assess if cytokine levels detected in platelet products can serve as indicators of acceptable platelet activation as a result of the WBC-reduction process. MATERIALS AND METHODS: We measured the levels of cytokines predominantly derived from WBCs [e.g. interleukin-8 (IL-8)] and platelets [e.g. regulated on activation, normal, T-cell expressed, and secreted (RANTES) and transforming growth factor-beta(1) (TGF-beta(1))] under the present experimental conditions in different WBC-reduced PCs, i.e. PCs prepared from three different WBC-reduction filters and control non-filtered PCs using pooled BCs from the same donors and three apheresis types. Supernatant plasma was collected at the beginning (day 1) and end (day 5) of the shelf life of each PC, and the cytokine content was determined using appropriate enzyme-linked immunosorbent assays (ELISAs). Process efficiency was assessed by platelet yield and residual WBC count. RESULTS: We found that products from the apheresis process involving a filtration step (Haemonetics MCS+) showed a lower cytokine content on both day 1 and day 5 in comparison with the fluidized bed (COBE Spectra) or elutriation (Amicus) processes. WBC reduction of BC-PCs of the same origin using three different filters showed comparable levels of cytokines on day 1 in all units. After storage for 5 days, the levels of IL-8 remained essentially unchanged in filtered BC-PCs but increased by more than threefold in control non-filtered BC-PCs, suggesting IL-8 release by residual WBCs present in the control PCs. The concentration of platelet-derived cytokines such as RANTES and TGF-beta(1), however, increased significantly in all filtered and control non-filtered PCs during the storage period. CONCLUSION: These results show that markers of cytokine release from both WBCs and platelets are useful indicators of the performance and efficacy of the WBC-reduction process and of platelet quality.     

Correlation of in vitro platelet quality measurements with in vivo platelet viability in human subjects

BACKGROUND AND OBJECTIVES: Changes in in vitro platelet quality parameters during platelet storage are associated with a decrease of in vivo platelet viability after platelet transfusion. Many attempts have been made to identify the most predictable in vitro parameters for in vivo performance. We used a riboflavin-based ultraviolet (UV) light treatment process designed to inactivate pathogens and white blood cell (WBC) contaminants in blood products as a model system in which to study the correlation of in vitro cell quality with in vivo viability. MATERIALS AND METHODS: Platelet products (n = 18) were collected by a standard Trima apheresis procedure and treated with one of three dose levels of UV light (0, 7.2 or 12.4 J/ml) in the presence of 50 microm riboflavin. Lactate production, glucose consumption and P-selectin expression, pH, pCO(2), pO(2), hypotonic shock response and swirl were measured during 5 days of platelet storage post-UV/RB treatment. Aliquots of these products were radiolabelled on day 5 of storage and were subsequently used to determine platelet recovery and survival time in autologous subjects. RESULTS: The responses of in vitro cell quality were observed to occur in a UV dose-dependent manner. Lactate production and pH were identified as the parameters most strongly correlated with platelet in vivo recovery, which ranged from 5 to 82%. The correlation coefficients (r) for lactate production and pH with in vivo recovery in human subjects were 0.9090 and 0.8831 with P-values of 0.007 and 0.031, respectively. Lactate production and pH were also found to be correlated with platelet survival time, with correlation coefficients of 0.8063 and 0.8384 (the P values were 0.01 and 0.001, respectively). CONCLUSIONS: Using conditions of riboflavin-based UV light treatment, lactate production and pH were identified as having the highest correlations with recovery and survival of radiolabelled platelets in healthy subjects.     

Storage of Platelet Concentrates from Overnight-Stored Blood and Overnight-Stored Buffy Coat: In vitro Studies

Platelet concentrates (PCs) were prepared from single buffy coats derived from fresh blood and from blood units stored overnight, as well as from buffy coats that were stored overnight. The platelet yield from overnight-stored buffy coats was similar to that of fresh blood or overnight-stored blood. PCs were stored at 20–24°C and on day 5 of storage, platelet aggregation with ADP was tested both at 37 and 25°C. Stored platelets aggregated better at 25°C than 37°C. The maximal aggregation (10 μM ADP) of stored platelets from overnight-stored buffy coats was 46±23% (n = 30), while that of stored platelets prepared either from fresh or overnight-stored blood was 27±21% (n = 29) and 22±15% (n = 29), respectively. Extracellular lactate dehydrogenase and ammonia levels, as well as elastase activity were similar in stored PCs of different origin. Our conclusion is that PCs prepared from overnight-stored buffy coat might also be suitable for storage and clinical use. In vivo studies are needed to confirm our findings.     

Storage of platelets in additive solutions: a multicentre study of the in vitro effects of potassium and magnesium

BACKGROUND AND OBJECTIVES: In a preliminary study, the presence of potassium and magnesium in a modified synthetic medium (PAS-III) was found to have a significant influence on platelet metabolism (using apheresis-derived, as well as buffy-coat-derived platelets) when compared with standard PAS-III. The differences included reduced glycolysis, as evidenced by lower consumption of glucose and lower production of lactate, but also better preservation of pH and hypotonic shock response reactivity. The results suggested that storage in modified PAS-III containing 20% plasma was comparable to storage in standard PAS-III containing 30% plasma. To confirm the preliminary results and to evaluate the effects of different preparation protocols, an international multicentre study, which included 11 different sites, was conducted. MATERIALS AND METHODS: Platelets from 30 pools of approximately 20 buffy coat (BC) units each and 24 pooled apheresis platelet units were aliquoted for storage in plasma (reference) or synthetic medium using either a specific additive solution (PAS-III) containing 30% plasma or a modification of PAS-III containing 5.0 mm potassium and 1.5 mm magnesium (PAS-IIIM) and either 30% or 20% plasma. Units were stored at room temperature with agitation for 7 days during which in vitro testing was carried out for biochemical, haematological and functional parameters. RESULTS: Storage of platelets in PAS-IIIM resulted in a reduction in the rate of glycolysis and better retention of pH and hypotonic shock response reactivity. Storage in PAS-IIIM containing 20% plasma appeared to result in the retention of in vitro properties, similar to those observed during storage in standard PAS-III containing 30% plasma. CONCLUSIONS: The results of this study confirm the preliminary results. Similar results were seen with platelets prepared by BC and apheresis methods, despite differences in equipment, the preparation technique and in the final platelet contents achieved in the platelet units. Storage of platelets in PAS-IIIM should be considered to improve platelet function and allow plasma reduction to 20%.     

Cold storage of pooled, buffy-coat-derived, leucoreduced platelets in plasma

Background  This study was designed to determine which in vitro assays would be most useful for studying the effects of cold storage on platelet concentrates and to establish an in vivo model for platelet recovery and survival.
Study Design and Methods  Paired, plasma-suspended, leucoreduced, buffy-coat-derived platelet concentrates were stored either at 22 or 4°C. Prior to storage and after 18 h, 5 days and 7 days, samples were taken and various assays were performed. On day 6, in vivo studies were carried out using a model system. Galactosylation of the platelets, prior to cold storage, was also tested.
Results  Hypotonic shock response, collagen-induced aggregation, RANTES and P-selectin binding site measurements demonstrated differences between platelets stored at 22 and 4°C. The glycocalicin assay was able to demonstrate microvesicle formation at 4°C. The in vivo model showed that there was at least a 50% decrease in recovery and survival when the platelets were stored in the cold. Galactosylation did not improve these results.
Conclusions  Several assays, both in vitro and in vivo , were able to detect differences in platelet-storage characteristics and in vivo recovery and survival in a model system. Galactosylation did not correct these cold-induced changes.     

Activation during preparation of therapeutic platelets affects deterioration during storage: a comparative flow cytometric study of different production methods

Three different separation methods, all using centrifugation, are routinely used to prepare therapeutic platelet concentrates from human donor blood. Platelet concentrates derived from platelet-rich plasma (PRP-PC), buffy coat (BC-PC) and apheresis (AP-PC) were investigated at the end of production, and over an 8 d storage period. Change in platelet surface markers were measured by flow cytometry, using fluorescein-conjugated antibodies to fibrinogen, P-selectin (CD62P), GPIIb–IIIa (CD41), GPIbα (CD42b) and GPV (CD42d), and fluorescein-conjugated Annexin V was used to measure expression of anionic phospholipid.
All concentrates showed some changes during preparation but PRP-PC underwent the greatest changes with significantly higher levels of P-selectin ( P  < 0.001) and bound Annexin V ( P  = 0.001) than AP-PC or BC-PC, and lower levels of GPIbα ( P  = 0.002) and GPV ( P  < 0.001). These changes were attributable to component separation rather than venesection. These markers all continued to change on storage with a strong positive correlation between the changes seen during production and those after 5 d storage. PRP-PC continued to show the greatest changes whereas BC-PC showed the least. Fibrinogen was bound to 40–50% of platelets in all preparations and this did not alter significantly on storage whereas total expression of GPIIb–IIIa remained unchanged throughout.
There was no evidence that the platelet surface changes were thrombin-mediated and leucocyte depletion of BP-PC by filtration had no effect on the changes. It is proposed that the deterioration of platelet concentrates during storage may be related to activation occurring during preparation. 'Whole blood' flow cytometry using a panel of fluorescein-labelled reagents provides an informative method for evaluating platelet concentrates.     

Microparticle and mitochondrial release during extended storage of different types of platelet concentrates

On activation, platelets release vesicles called microparticles (MPs). MPs are heterogeneous with regard to the presence or absence of mitochondria. We quantified MPs in platelet concentrates (PCs) taking their mitochondrial content into account. Platelet-rich plasma (PRP), buffy coat (BC) and apheresis (AP) PCs were tested through 7 days of storage. A combination of flow cytometry and spanning-tree progression analysis of density-normalized events (SPADE) was used to determine MP and mitochondrial release during storage. All the PC biochemical parameters complied with transfusion standards at all times. Platelet activation markers increased during storage and were higher for PRP than other types of PCs. Concentrations of MPs and extracellular mitochondria interpreted by SPADE algorithm were significantly higher in PRP than other in PCs and were stable throughout storage. The mode of preparation, rather than storage duration, impacts the release of MPs and mitochondria in PCs.     

In vitro and in vivo effects of potassium and magnesium on storage up to 7 days of apheresis platelet concentrates in platelet additive solution

BACKGROUND AND OBJECTIVE: Prolonged storage of platelets up to 7 days provides improved availability, logistical management and decreased wastage. Beside methods of bacterial detection, addition of magnesium and potassium to the platelet storage solution (SSP+) may further improve the quality of platelets with extended storage. MATERIALS AND METHODS: Apheresis platelets from 10 donors were divided and stored in two different platelet additive solutions (PAS) (Intersol and SSP+) for a paired comparison. A variety of in vitro platelet function and metabolic assays were performed both on day 1 and after 7 days of storage. For in vivo study, platelets were labelled with either (111)Indium or (51)Chromium after 7 days of storage and were injected into the corresponding donor. Serial blood samples were drawn for recovery and survival measurements. RESULTS: In vitro parameters for SSP+ showed significantly reduced glycolysis (lower glucose consumption and decreased production of lactate), a higher hypotonic shock response (HSR) and the extent of shape change reactivity and a lower degree of platelet activation by means of RANTES (regulated on activation, normal, T cell-expressed, and secreted), CD62p and CD63 expression. Platelet recovery on day 7 was higher for Intersol as compared to SSP+, 65 +/- 11 vs. 53 +/- 13% (P = 0.023), and survival showed no difference 4.2 +/- 1.9 vs. 3.6 +/- 1.4 days. CONCLUSION: In vitro characteristics of platelets stored in PAS with addition of potassium and magnesium indicated higher quality, but this could not be verified by the in vivo parameters by means of recovery and survival.     

In vitro function of platelet concentrates prepared after filtration of whole blood or buffy coat pools

BACKGROUND/METHOD: Data on the quality of platelet concentrates (PC) produced by the buffy coat method and stored beyond 5 days in plasma are limited. We therefore evaluated the quality of PCs prepared by leucocyte depletion of whole blood (Terumo WBSP, n = 10) or a buffy coat pool (Pall Autostop, n = 10), and stored for 7 days in plasma by assessing platelet parameters and markers of platelet activation. RESULTS: In both types of PC, levels of glucose decreased during storage but were not totally depleted (> 11 mM on day 7). In contrast, lactate levels increased on storage and was consistently < 20 mM throughout, with pH maintained at > 6.8 in all units. Hypotonic shock response scores were > 47% in all units at day 7. On day 1, markers of platelet activation were significantly higher in WBSP PC, but by day 7 were similar for percentage CD63+ and CD62P + (40%) with levels of platelet microparticles and annexin V binding two-fold higher in WBSP. The expression of CD61 did not alter during storage and the percentage of platelets expressing CD42b was > 88% in all units on day 7. RANTES (Regulated on activation, normal, T-cell expressed and secreted) and TGFbeta released from platelets by day 7 was < 800 ng/ml and 90 ng/ml, respectively. C3a(desarg) increased throughout storage in both types of PC, but without a commensurate increase in the terminal complex SC5b-9 or activation of factor XII. CONCLUSION: Our data indicates that the in vitro characteristics of PCs prepared using these methods is maintained over storage for 7 days in plasma and is not associated with significant deterioration of platelet function. ONE SENTENCE SUMMARY: In vitro function of platelet concentrates prepared by either filtration of whole blood, or pooled buffy coats.     

Enhanced circulatory parameters of human platelets cryopreserved with second-messenger effectors: an in vivo study of 16 volunteer platelet donors

Platelet transfusion represents an important component of the therapy for thrombocytopenic patients. Prolonged storage capabilities for platelets would alleviate many problems associated with blood banking. Unfortunately, current cryopreservation methods are complex to implement and result in loss of cell number and functional activity. Previous in vitro studies have shown that the use of ThromboSolTM, a platelet-stabilizing formulation, in the cryopreservation of platelets results in significant retention of cell number and in vitro functional activities in addition to reducing the DMSO requirement to only 2%. We evaluated the in vivo circulatory parameters of platelets cryopreserved with ThromboSol. Single donor platelet units were obtained from healthy volunteers (n = 16); the units were then split and cryopreserved with either ThromboSol and 2% DMSO or 6% DMSO alone. Following storage at -80 degrees C for 7-10 d the samples were thawed, washed and radiolabelled with either 51Cr or 111In. The paired samples were then mixed and reinfused into the autologous volunteer. At various time intervals following transfusion a blood sample was drawn and the quantity of circulating labelled platelets was determined. The percent recovery and survival time was determined by multiple-hit analysis. The ThromboSol-treated platelets, as compared to the 6% DMSO-treated platelets, displayed statistically higher percent recovery (40.2% v 28.8%) and survival time (166.3 h v 152.1 h). These results demonstrated that platelets cryopreserved with ThromboSol displayed superior in vitro and in vivo characteristics as compared to the standard 6% DMSO method. The use of ThromboSol allowed for a 3-fold reduction in the DMSO concentration in conjunction with a 40% increase in circulating cell number and normal survival times.     

In vitro assessment of apheresis and pooled buffy coat platelet components suspended in plasma and SSP+ photochemically treated with amotosalen and UVA for pathogen inactivation (INTERCEPT Blood System™)

Background and Objectives INTERCEPT Blood System? is a pathogen inactivation system for blood components. The initial approval required a platelet component to be suspended in a combination of plasma and Platelet additive Solution/PAS‐III. Improved platelet storage has been reported with Mg++ and K+ supplementation (PAS‐IIIM). This study validated the use of INTERCEPT?/PAS‐IIIM for apheresis and pooled buffy‐coat platelet components. Materials and Methods The platelet dose and pH throughout 5 days of storage met the European and French requirements for quality standards. Results and Conclusion Additional metabolic and activation assessments of the treated platelets confirmed the previously reported superiority of PAS‐IIIM over PAS‐III, but extended it to the INTERCEPT? process.     

Functional characteristics of S-59 photochemically treated platelet concentrates derived from buffy coats

BACKGROUND: A photochemical treatment (PCT) process for inactivation of infectious pathogens and leukocytes has been developed and evaluated using single-donor platelet concentrates. This study assessed the application of PCT to platelets prepared from pooled buffy coats. In this study, in vitro functional characteristics of PCT platelets were compared to control platelets prepared from pooled buffy coats using the approved platelet-additive solution T-Sol((R)). Platelets in platelet PAS III additive solution without PCT were evaluated as well. PCT also included the use of a psoralen (S-59) reduction device (SRD). MATERIALS AND METHODS: Four types of platelet concentrates were compared: (1) platelet concentrate in plasma/T-Sol; (2) platelet concentrate in plasma/PAS III; (3) platelet concentrate in plasma/PAS III, PCT, 9 h SRD and (4) platelet concentrate in plasma/PAS III, PCT, 16 h SRD. PCT occurred on the day after whole-blood collection. In vitro assay parameters included: pH, pO(2), pCO(2), HCO(-)(3), platelet count, mean platelet volume, plasma glucose, plasma lactate, total ATP, expression of p-selectin, hypotonic shock response and electron microscopy. RESULTS: The results indicate that PCT is compatible with platelet concentrates prepared from pooled buffy coats for up to 7 days of storage. CONCLUSION: The PCT process resulted in acceptable in vitro platelet functional characteristics and is currently in clinical trials to evaluate the haemostatic efficacy of PCT platelets in thrombocytopenic patients requiring multiple platelet transfusions.     

Evaluation of Apheresis Platelet Concentrates Collected with a Reduced (30-ml) Collection Chamber with Resuspension and Storage in a Synthetic Medium

Recently, the CS-3000® Plus Blood Cell Separator with the TNX-6 platelet separation chamber insert has been furnished with a small-volume (30-ml) collection chamber. In this study, a platelet synthetic medium containing glucose and bicarbonate (PSM) was used for resuspension and storage of this highly concentrated platelet product. Eighteen donors participated in a paired study design where each participant donated platelets on two occasions, once following collection in a standard chamber with resuspension and storage in plasma and once following collection in the new chamber with resuspension and storage in PSM. Substantially higher total platelet counts were obtained using platelets collected in the small chamber and stored in PSM as compared to control (4.4±0.9times10¹¹ vs. 3.5±0.9times10¹¹ platelets, p<0.01 by paired t test). After 5 days of storage, PSM-stored platelets demonstrated higher ATP levels, less lactate dehydrogenase in the supernatant and increased lactate production with resulting lower pH at day 5 of storage (6.94±0.15 vs. 7.08±0.09, p<0.05). There were no statistically significant differences of the survival by multiple-hit estimation of PSM-stored as compared to plasma-stored platelets as determined by ¹¹¹In labeling and infusion. A slight decrease in the initial percent recovery with the additive-suspended as compared to suspended plasma cells was noted: 50±8 versus 54±9%, respectively (p<0.05). In conclusion, the CS-3000 Plus/TNX-6 apheresis system with a new reduced-volume collection chamber and an additive solution provides a plasma-poor and highly concentrated platelet product with satisfactory in vivo viability and in vitro functional characteristics after 5 days of storage.