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1.  Comparison of plateletpheresis on three continuous flow cell separators 
Platelet concentrate (PC) remains one of the most important support measures in thrombocytopenic patients. An efficient cell separator is a prerequisite for an optimally functioning apheresis setup. Donor blood count may undergo a temporary reduction after the procedure.
The aim was to find the extent of reduction in donor blood count (hemoglobin, hematocrit, white blood cell, and platelet) after plateletpheresis and to evaluate the cell separator for collection efficiency, processing time, and leukoreduction.
Study Design and Methods:
Two hundred and thirty seven procedures performed on the Amicus (N = 121), Fenwal CS-3000 Plus (N = 50) and Cobe spectra (N = 66) in a one year period were evaluated. The procedures performed on the continuous flow centrifugation (CFC) cell separators and donor blood counts (pre and post donation) done were included in the study.
The percent reduction in hemoglobin (HB), hematocrit (HCT), white blood cell (WBC) and platelet count ((PLT ct) was 2.9, 3.1, 9, 30.7 (Mean, N = 237) respectively after the procedure. The post donation PLT ct reduced to < 100×109/L (range 80-100) in five donors (N = 5/237, Amicus). The pre donation PLT ct in them was 150-200×109/L. Collection efficiency (percent) of Amicus (79.3) was better as compared to the other two machines (CS: 62.5, Cobe: 57.5). PC collected on Cobe spectra had <1×106 WBC. The donor pre donation PLT levels had a positive correlation to the product PLT yield (r = 0.30, P = 0.000).
Monitoring donor blood counts helps to avoid pheresis induced adverse events. A cautious approach is necessary in donors whose pre donation PLT ct is 150-200×109/L. The main variable in PLT yield is donor PLT ct (pre donation). High collection efficiency is a direct measure of an optimally functioning cell separator.
PMCID: PMC2920476  PMID: 20808650
Continuous flow cell separator; donor blood count; plateletpheresis; platelet yield; Blood donor; apheresis
2.  Platelet Recruitment during Multiple Donor Platelet Apheresis Differs between Cell Separators 
Recruitment of platelets (PLT) during donor PLT apheresis may facilitate the harvest of multiple units within a single donation.
We compared two PLT apheresis procedures (Amicus and Trima Accel) in a prospective, randomized, paired cross-over study in 60 donors. The 120 donations were compared for depletion of circulating PLT in the donors, PLT yields and PLT recruitment. A recruitment was defined as ratio of total PLT yield and donor PLT depletion > 1.
Despite comparable differences of pre- and post-apheresis PLT counts (87 × 109/l in Trima Accel vs. 92 × 109/l in Amicus, p = 0.383), PLT yields were higher with Trima Accel (7.48 × 1011 vs. 6.06 × 1011, p < 0.001), corresponding to a higher PLT recruitment (1.90 vs. 1.42, p < 0.001). We observed a different increase of WBC counts after aphereses, which was more pronounced with Trima Accel than with Amicus (1.30 × 109/l vs. 0.46 × 109/l, p < 0.001).
Both procedures induced PLT recruitment. This was higher in Trima Accel, contributing to a higher yield in spite of a comparable depletion of circulating PLT in the donors. This recruitment facilitates the harvest of multiple units within a single donation and seems to be influenced by the procedure utilized. The different increases of circulating donor white blood cells after donation need further investigation.
PMCID: PMC3128152  PMID: 21760764
Platelet apheresis; Platelet recruitment; Cell separator
3.  Pre- and post- donation haematological values in healthy donors undergoing plateletpheresis with five different systems 
Blood Transfusion  2009;7(3):188-192.
Although automated cell separators have undergone a lot of technical refinements, attention has been focused more on the quality of platelet concentrates than on donor safety. We planned this prospective study to observe the effects of automated plateletpheresis on normal haematological values of healthy donors and to determine whether the haematological alterations had any clinical consequences.
Study design and methods
The study was conducted on 457 healthy, first-time plateletpheresis donors over a period of 26 months. The plateletpheresis procedures were performed using five different cell separators and various pre- and post-donation haematological values such as haemoglobin concentration (Hb), haematocrit (Hct), platelet and white blood cell (WBC) counts, mean platelet volume and platelet distribution width were measured in all donors.
We observed that the Hb, Hct, platelet and WBC counts decreased significantly in the donors (p<0.01) after each procedure, without there being significant changes in mean platelet volume or platelet distribution width. The decreases in Hb and Hct were significantly greater with the CS 3000 and Amicus machines, while the decreases in platelet and WBC counts were significantly greater with the CS 3000 and Fresenius separators.
Although a significant drop in complete blood count was observed in all donors, none manifested features of thrombocytopenia or anaemia. Nevertheless, more prospective studies on this aspect are required in order to establish guidelines for donor safety in apheresis and also to help in assessing donor suitability, especially given the present trend of double product apheresis collections.
PMCID: PMC2719270  PMID: 19657482
Plateletpheresis; haematological values; cell separator; donor safety; platelet count
4.  Adverse events associated with apheresis procedures: Incidence and relative frequency 
Apheresis procedures [Plateletpheresis, Plasmapheresis/ Therapeutic Plasma Exchange (TPE), & Peripheral Blood Stem Cell Collection (PBSC)] are usually well tolerated. Occasionally, Adverse Events (AEs) of variable severity may occur during or after the procedure. AEs that occur in Donors/Patients are divided into local reactions and systemic reactions.
Materials and Methods:
A total of 3,367 apheresis procedures were performed, out of which 3,120 were plateletpheresis procedures, and out of which 1,401 were on Baxter CS 3000 & 1,719 were on Haemonetics MCS+ cell separators. Rest of 247 TPE & PBSC procedures were done on Haemonetics MCS+ cell separators.
90 AEs were reported in relation to the 3,367 procedures. Out of 90 AEs, 85 AEs (94%) were associated with plateletpheresis (n = 3,120) and 05 AEs (06%) with TPE & PBSC (n = 247). The rate of vascular injury (VI), Citrate reaction (CR), and Presyncopal/Syncopal (PS/S) in plateletpheresis was 1.6% (52/3,120), 0.96% (30/3,120), and 0.096% (03/3,120), respectively. The rate of CR in TPE and PBSC was 1.23% (02/162) and 2.3% (02/85), respectively. The rate of PS/S in PBSC was 1.17% (01/85). AEs for Plateletpheresis, TPE & PBSC were 2.7% (85/3,120), 1.23% (02/162), and 3.5% (03/85), respectively. VI, CR, and PS/S were mostly of mild intensity. Both cell separators were equally safe, when AEs associated with plateletpheresis were compared with each other; 2.8% on CS 3000 & 2.6% on MCS+.
Apheresis procedures performed on cell separators are safe, with a low incidence of significant AEs. No significant difference was noted in AEs among the two cell separators studied.
PMCID: PMC3613659  PMID: 23559763
Adverse events; citrate reaction; peripheral blood stem cell; presyncopal/syncopal; therapeutic plasma exchange; vascular injury
5.  Probiotic-associated high-titer anti-B in a group A platelet donor as a cause of severe hemolytic transfusion reactions 
Transfusion  2009;49(9):1845-1849.
Hemolytic transfusion reactions (HTRs) can occur with transfusion of platelets (PLTs) containing ABO-incompatible plasma. Reported cases have involved group O donors. Two cases of PLT-mediated HTRs associated with the same group A plateletpheresis component, collected from a donor taking high doses of probiotics are reported.
Case 1 was a 40-year-old 69-kg group B stem cell transplant patient who received one-half of a group A plateletpheresis component. Severe back pain occurred 10 minutes into the transfusion, accompanied by anemia and hyperbilirubinemia. Case 2 was a 5-year-old 26-kg group B male with aplastic anemia who received the other half of the same plateletpheresis component, volume reduced to 37 mL. Syncope occurred immediately after the transfusion, with laboratory evidence of hemolysis a few hours later.
Serologic investigation of posttransfusion samples from both patients revealed positive direct anti-globulin tests: C3d only for Case 1 and immunoglobulin (Ig)G and C3d for Case 2; the eluates contained anti-B. The group A donor’s anti-B titer was 16,384 at saline and IgG phases. Donor lookback revealed that the donor had donated 134 apheresis PLTs over many years. For 3 years, he had intermittently taken probiotics; 3 weeks before the index donation, he began taking three tablets of probiotics every day. Lookback of prior group B recipients uncovered a case of acute hemolysis that was not recognized at the time. The solubilized probiotic inhibited anti-B in vitro.
Non–group O PLT donors can have high-titer anti-A or anti-B that might mediate HTRs, and probiotic ingestion in blood donors represents a novel mechanism of stimulating high-titer anti-B.
PMCID: PMC3421026  PMID: 19453987
6.  Adverse reactions in blood and apheresis donors: experience from two Italian transfusion centres 
Blood Transfusion  2009;7(1):35-38.
Blood and apheresis donations are widely considered to be safe with a low incidence of adverse reactions and injuries; however, data reported in the medical literature on the prevalence of adverse events in donors and studies on the predictive risk factors for donor reactions are limited and contradictory.
From January 2002 to December 2006 we recorded every adverse reaction verified during 240,596 consecutive blood and apheresis donations (183,855 homologous whole blood donations, 6,669 autologous whole blood donations, 38,647 plasmapheresis, 2,641 plateletpheresis and 8,784 multicomponent donations) at the Italian Transfusion Centres of Verona and Ragusa,.
Using a special, pre-arranged form within the quality system, a total of 686 adverse reactions (related to 0.28% of all donations) were recorded. Vasovagal reactions, mostly of mild intensity, were the most commonly observed adverse reactions, with a frequency of 0.20% (487/ 240,596). The frequency of the vasovagal reactions varied according to the different types of donation, being 0.19% (346/183,855) for homologous whole blood donations, 0.24% (16/6,669) for autologous whole blood donations, 0.16% (63/38,647) for plasmapheresis, 0.68% (18/2,641) for plateletpheresis and 0.49 (43/8,784) for multicomponent donations. Citrate toxicity was reported in 0.38% (189/50,072) of apheresis donations. Severe adverse reactions were very rare, as they occurred in 0.004% of the donations (10/240,596).
In conclusion, the results of our 5-year survey document that apheresis and blood donation are safe procedures for the donor with a low incidence of adverse reactions; the adverse reactions that did occur were mostly mild and resolved rapidly.
PMCID: PMC2652234  PMID: 19290078
blood donation; apheresis; adverse events
7.  Initial Values of Donor Hematocrit and Efficiency of Plateletpheresis 
Acta Informatica Medica  2013;21(2):116-119.
The collection of platelets by apheresis is considered as a very great progress in transfusion medicine. A larger yield (total number of collected platelets) is obtained if the donor has a greater number of initial platelets and if the separation is done in a shorter time. One of the parameters is also the efficiency of the platelet collection (expressed in percentage) on the value of which different factors may have direct or indirect influence.
To calculate the efficiency of platelet collection with the separator Fenval Baxter AMICUS and to compare the efficiency of platelet collection with this separator in relation to the initial value of donor hematocrit.
Donors and Methods
The donors who participated in this study were divided into groups according to the value of the donor’s ‘hematocrit before separation. Group C consisted of donors whose initial value of the hematocrit was lower or equal to 46%. Group D consisted of donors whose initial value of the hematocrit was higher than 46%. The process was carried out on Fenval Baxter AMICUS. The expected efficiency of the collection was obtained by dividing the total number of collected cells by the expected total number of processed cells, i.e. the total number of cells passed through the equipment.
In the 258 separations which satisfied the fixed criteria were men in 226 cases (87.6%) and women in 32 (12.4%). There is a statically significant difference in the platelet value between the groups and this value is higher in group D than in group C. The average value of platelets before separation was 46.66. The range of minimal and maximal value is from 38.8 to 52.4 ±2.78. The initial value of hematocrits of the donor does not intervene in the length of the separation, but it has a significant effect on the efficiency of the platelet collection. Increases in the number of hematocrits significantly decrease the efficiency of platelet collection. In practice it means that we can base on this fact make a better selection of donors. In this kind selection, one should prefer a donor with a higher number of initial platelets and lower levels of hematocrits. In that way we can collect a more important yield, have a shorter length of separation and increase the efficiency of platelet collection. Its advantage is as well medical because of a more important yield but also financial because of the decrease of the length of the separation and the increase in efficiency. Key words: value of hematocrits, donors, apheresis, platelets, efficiency.
PMCID: PMC3766539  PMID: 24058252
value of hematocrits; donors; apheresis; platelets; efficiency
8.  Evaluation of White Blood Cell- and Platelet-Derived Cytokine Accumulation in MIRASOL-PRT-Treated Platelets 
Soluble mediators in platelet concentrates (PCs) released from contaminating white blood cells (WBCs) and platelets (PLTs) themselves are supposed to promote allergic and non-hemolytic febrile transfusion reactions in the recipient. Pathogen reduction technologies (PRTs) prevent replication and proliferation of pathogens as well as of WBCs, and may reduce cytokine accumulation in PCs during storage and prevent adverse events after PLT transfusion. On the other hand, such treatments may also lead to increased cytokine production by stimulation of WBCs or PLTs due to the photochemical or photodynamical process itself.
Material and Methods
12 triple-dose PLT apheresis collections were leukoreduced by the process-controlled leukoreduction system of the Trima Accel machine and split into 3 units undergoing Mirasol-PRT treatment (M) or gamma irradiation (X) or remaining untreated (C). During storage for up to 7 days, PLT activation, WBC-derived Th-1/2, and inflammatory as well as PLT-derived cytokines were measured by cytometric bead array and enzymelinked immunosorbent assay, respectively.
Independent of treatment, all PLT products exhibited low levels of WBC-associated cytokines near or below assay detection limits. WBC-associated cytokines were not elevated by Mirasol-PRT treatment. PLT-derived cytokines were detected at higher levels and increased significantly during storage in all units. Most likely due to higher PLT activation, M units showed significantly higher levels of PLT-derived cytokines compared to untreated and gamma-irradiated units on day 5 of storage.
In all PCs, PLTs themselves were the main source of cytokine release. Mirasol-PRT treatment was associated with a significantly increased PLT activation and accumulation of PLT-derived cytokines during storage, without affecting WBC-derived cytokines relative to controls.
PMCID: PMC2928824  PMID: 20823992
Pathogen reduction; Mirasol-PRT; Ccytokines; Transfusion reaction
9.  Effect of storage on levels of nitric oxide metabolites in platelet preparations 
Transfusion  2012;53(3):637-644.
Nitric oxide (NO), a potent signaling molecule, is known to inhibit platelet function in vivo. We investigated how the levels of NO and its metabolites change during routine platelet storage. We also tested whether the material of platelet storage containers affects nitrite content since many plastic materials are known to contain and release nitrite.
Study design and methods
For nitrite and nitrate measurement, leukoreduced apheresis platelets (PLT) and concurrent plasma (CP) were collected from healthy donors using the Trima Accel. Sixty mL aliquots of PLT or CP were stored in CLX or PL120 Teflon containers at 20–24°C with agitation and daily samples were processed to yield PLT pellet and supernatant. In a separate experiment, PLT was stored in PL120 Teflon to measure NO generation using electron paramagnetic resonance (EPR).
Nitrite level increased markedly in both PLT supernatant and CP stored in CLX containers at a rate of 58 nM/day and 31 nM/day respectively. However, there was a decrease in nitrite level in PLT stored in PL120 Teflon containers. Nitrite was found to leach from CLX containers and this appears to compensate for nitrite consumption in these preparations. Nitrate level did not significantly change during storage.
Platelets stored at 20–24°C maintain measurable levels of nitrite and nitrate. Nitrite decline in non-leachable Teflon containers in contrast to increases in CLX containers which leach nitrite, suggests that it is consumed by platelets, residual leukocytes or erythrocytes. These results suggest NO-related metabolic changes occur in platelet units during storage.
PMCID: PMC3548060  PMID: 22804724
nitric oxide; nitrite; platelet storage; transfusion
10.  Evaluation of low red blood cell mean corpuscular volume in an apheresis donor population 
Transfusion  2009;49(9):1971-1976.
Apheresis donors are routinely evaluated with a complete blood count (CBC). Low red blood cell mean corpuscular volume (MCV) values (<80 fL) in the presence of an acceptable hemoglobin (Hb; ≥12.5 g/dL) could be due to iron deficiency or hemoglobinopathy. The etiology of a low MCV in a healthy apheresis donor population was assessed.
Predonation samples for CBC were obtained from 1162 consecutive apheresis donors. Donors with a MCV of less than 80 fL were evaluated by CBC, iron studies (ferritin, serum iron, transferrin, percentage of transferrin saturation), and hemoglobin (Hb) electrophoresis. Iron deficiency was defined as a ferritin value below the reference range. Beta chain Hb variants were determined by Hb electrophoresis. Alpha thalassemia trait was presumed if the red blood cell (RBC) count was elevated, no variant Hbs were detected, and the iron studies were within normal ranges.
In a 19-month period, 33 of 1162 apheresis donors had low MCV values. Iron deficiency was present in 64%; 49% had isolated iron deficiency and 15% had iron deficiency plus hemoglobinopathy. Hemoglobinopathy without concomitant iron deficiency was found in the remaining 36%.
Iron deficiency is present in the majority of apheresis donors with repeatedly low MCV values and Hb levels of 12.5 g/dL or more. Hemoglobinopathy is also commonly present but may not be easily recognized in the setting of iron deficiency. The MCV is a useful screening tool to detect iron deficiency and hemoglobinopathy. Low MCV values should be investigated to determine if iron replacement therapy is indicated.
PMCID: PMC3421029  PMID: 19453988
11.  Emergency therapeutic leukapheresis in a case of acute myeloid leukemia M5 
Cell separators in India are routinely used for plateletpheresis, peripheral blood stem cell collections and therapeutic plasma exchange. Therapeutic leukapheresis, particularly as an emergency procedure, has been uncommonly performed and reported. Here, a case of a 53-year-old male, diagnosed with acute myeloid leukemia subtype M5 (AML M5) with hyperleukocytosis, who underwent emergency leukaphereis, is reported. After two procedures, there was a decrease of WBC count by 85%, which enabled cytotoxic therapy to be initiated.
PMCID: PMC2798757  PMID: 20041073
Emergency; hyperleukocytosis; leukapheresis
12.  A General Change of the Platelet Transfusion Policy from Apheresis Platelet Concentrates to Pooled Platelet Concentrates is Associated with a Sharp Increase in Donor Exposure and Infection Rates 
We compare the actual with the potential donor exposure and possible infection rates in the Hanover Medical School (MHH) platelet (PLT) transfusion recipients if the current MHH standard of apheresis PLT concentrate (A-PC) supply would be replaced by a pooled PLT concentrate (P-PC) transfusion regimen.
Donors, Patients, and Methods
The electronic records of the MHH Institute of Transfusion Medicine and the MHH Department of Medical Controlling were evaluated to assess the development of PLT needs and supply at MHH from 2003–2006. For 2006, we evaluated all PLT transfusion recipients with respect to their overall transfusion needs, classified them for low and high PLT transfusion needs, and related them to the diagnostic groups that underlie their PLT demands. We assumed a P-PC preparation procedure using 4 whole blood-derived buffy coats for all calculations for potential donor exposure. To predict the possible infection rates of an unrecognized viral infection with low prevalence in the general population to A-PC or to P-PC recipients and the influence of neutralizing agent specific antibodies (NAB), we established a mathematical contamination/infection model based on the current PLT transfusion mode and data about GBV-C virus infection among Hanover blood donors.
From 2003 to 2006, the 1,300–1,400 persons comprising MHH apheresis donor pool covered a 36% increase in PC transfusions. The exclusive use of P-PCs instead of A-PC would require a total of 36,240–49,276 whole blood donations to meet MHH demands, corresponding to a more than 1 log step increase in donor exposure. For individual hematological patients, the change to P-PCs would imply an 80–125%, for individual surgical patients a 40–50% higher donor exposure. Our infection model revealed an approximately 4 times higher infection.
A change to P-PC would imply a more than one log step higher donor exposure, and an unrecognized infection with a prevalence around 1% leads to an up to 4 times higher infection rate. A general change in the PC transfusion policy that favors P-PCs is dangerous and must be avoided.
PMCID: PMC3076344  PMID: 21512637
Donor exposure; Apheresis platelet concentrates; Pooled platelet concentrates; Infection rates from apheresis platelet concentrates; Infection rates from pooled platelet concentrates
13.  A single apheresis procedure in the donor may be enough to complete an allograft using the “Mexican Method” of non-ablative allografting 
Blood Transfusion  2009;7(2):127-131.
Since 1999, in Mexico we have been using a regimen to conduct allografts that involves non-myeloablative conditioning and peripheral blood stem cells (PBSC) and have introduced some changes with the main goal of decreasing the cost of the procedure.
Materials and methods
We analysed the salient apheresis features of a group of 175 allogeneic peripheral blood stem cell transplants conducted in two institutions in a 7-year period. The grafts were conducted using the “Mexican” non-myelo ablative conditioning regimen employing oral busulphan, i.v. cyclophosphamide and i.v. fludarabine. In all instances, the apheresis machine employed was the Baxter CS3000 Plus and donors were mobilised with filgrastim. The apheresis procedures were performed on days 0, +1 and +2, the end-point of collection being 5,000 mL of blood/m2 in each procedure. Three apheresis sessions were planned but the number was adjusted according to the cell yield.
The final number of allografted CD34 cells ranged between 0.5 and 25.4 × 106/Kg of the recipient’s body weight (median, 5.2 × 106/Kg). One to three apheresis procedures were needed to obtain a product containing more than 0.5 × 106 CD34 cells/Kg of the recipient, the median being two procedures; in 72 cases (41%) a single apheresis procedure was sufficient to obtain the target number of CD34 cells. The volumes of apheresis ranged between 50 and 600 mL (median, 400 mL).
Since the median cost of each apheresis procedure is 900 USD, the fact that two apheresis procedures was spared in 72 cases and one apheresis was spared in another 65 cases, led to a total saving of approximately 188,100 USD. It can be concluded that, in many cases, allogeneic transplants can be completed with a single apheresis session and that there are considerable financial benefits from this practice.
PMCID: PMC2689067  PMID: 19503634
apheresis; allografts; costs saving; transplantation
14.  Quality assessment of platelet concentrates prepared by platelet rich plasma-platelet concentrate, buffy coat poor-platelet concentrate (BC-PC) and apheresis-PC methods 
Platelet rich plasma-platelet concentrate (PRP-PC), buffy coat poor-platelet concentrate (BC-PC), and apheresis-PC were prepared and their quality parameters were assessed.
Study Design:
In this study, the following platelet products were prepared: from random donor platelets (i) platelet rich plasma - platelet concentrate (PRP-PC), and (ii) buffy coat poor-platelet concentrate (BC-PC) and (iii) single donor platelets (apheresis-PC) by different methods. Their quality was assessed using the following parameters: swirling, volume of the platelet concentrate, platelet count, WBC count and pH.
A total of 146 platelet concentrates (64 of PRP-PC, 62 of BC-PC and 20 of apheresis-PC) were enrolled in this study. The mean volume of PRP-PC, BC-PC and apheresis-PC was 62.30±22.68 ml, 68.81±22.95 ml and 214.05±9.91 ml and ranged from 22-135 ml, 32-133 ml and 200-251 ml respectively. The mean platelet count of PRP-PC, BC-PC and apheresis-PC was 7.6±2.97 × 1010/unit, 7.3±2.98 × 1010/unit and 4.13±1.32 × 1011/unit and ranged from 3.2 –16.2 × 1010/unit, 0.6-16.4 × 1010/unit and 1.22-8.9 × 1011/unit respectively. The mean WBC count in PRP-PC (n = 10), BC-PC (n = 10) and apheresis-PC (n = 6) units was 4.05±0.48 × 107/unit, 2.08±0.39 × 107/unit and 4.8±0.8 × 106/unit and ranged from 3.4 -4.77 × 107/unit, 1.6-2.7 × 107/unit and 3.2 – 5.2 × 106/unit respectively. A total of 26 units were analyzed for pH changes. Out of these units, 10 each were PRP-PC and BC-PC and 6 units were apheresis-PC. Their mean pH was 6.7±0.26 (mean±SD) and ranged from 6.5 – 7.0 and no difference was observed among all three types of platelet concentrate.
PRP-PC and BC-PC units were comparable in terms of swirling, platelet count per unit and pH. As expected, we found WBC contamination to be less in BC-PC than PRP-PC units. Variation in volume was more in BC-PC than PRP-PC units and this suggests that further standardization is required for preparation of BC-PC. As compared to the above two platelet concentrates, all the units of apheresis-PC fulfilled the desired quality control criteria of volume. Apheresis-PC units showed better swirling and platelet count than PRP-PCs and BC-PCs. All the platelet concentrates units had pH well above the recommended norm.
PMCID: PMC2920479  PMID: 20808653
Corrected count increment; buffy coat poor-platelet concentrate; percentage recovery; platelet concentrate; platelet rich plasma-platelet concentrate; random donor platelet; single donor platelets
15.  The determinants of granulocyte yield in 1198 granulocyte concentrates collected from unrelated volunteer donors mobilized with dexamethasone and granulocyte–colony-stimulating factor: a 13-year experience 
Transfusion  2008;49(3):421-426.
The combination of granulocyte–colony-stimulating factor (G-CSF [filgrastim]) and dexamethasone (G-CSF/dex) is an effective granulocyte mobilization regimen, but the variables that affect donor neutrophil response and granulocyte collection yield are not well characterized.
A computerized database containing records of 1198 granulocyte collections from 137 unrelated volunteer apheresis donors during a 13-year period was retrospectively analyzed. Donors were categorized by age, sex, and cumulative number of granulocyte donations. Complete blood counts at baseline and after G-CSF/dex stimulation were recorded. The outcome variables include the pre-procedure absolute neutrophil count (preANC), which reflects G-CSF/dex stimulation, and the granulocyte product yield per liter processed (BagGranYield/L).
Higher baseline ANC and platelet (PLT) counts were significantly associated with higher preANC while a larger number of prior granulocytapheresis procedures was associated with lower preANC. Total filgrastim dose (used in weight-based dosing) did not significantly impact preANC or the granulocyte yield; weight-based dosing at 5 μg per kg and a uniform 480-μg dose produced equivalent preANC. PreANC and weight were the key determinants of granulocyte yield (BagGranYield/L).
Apheresis donors with higher baseline PLT counts and ANCs have higher ANCs after G-CSF/dex stimulation; donor age, weight, and sex do not have a significant impact. A uniform G-CSF dose of 480 μg is as effective as weight-based dosing at 5 μg per kg. Donor ANC monitoring should be considered after serial granulocytapheresis procedures.
PMCID: PMC3421027  PMID: 19040597
16.  Gravity sedimentation of granulocytapheresis concentrates with hydroxyethyl starch efficiently removes red blood cells and retains neutrophils 
Transfusion  2010;50(6):1203-1209.
Transfusion of granulocytapheresis concentrates can be limited by the volume of incompatible donor red blood cells (RBCs) in the component. Efficient reduction of RBCs in granulocyte units would result in safe transfusion of RBC-incompatible units.
Granulocyte concentrates were collected by continuous-flow apheresis from granulocyte–colony-stimulating factor (G-CSF) and dexamethasone-stimulated volunteer donors, with 6% hydroxyethyl starch (HES) added continuously during apheresis as a RBC sedimenting agent to enhance granulocyte collection efficiency. After collection, the component was placed in a plasma extractor for 4 hours. A sharp line of demarcation between the starch-sedimented RBCs and the granulocyte-rich supernatant developed, and the supernatant was transferred to a sterilely docked transfer pack. RBC reduction and white blood cell recovery were determined.
Gravity sedimentation was performed on 165 granulocyte concentrates. Mean sedimentation time was 267 minutes (range, 150–440 min). RBC depletion was 92% (range, 71%–99%) with mean residual RBC content of 3.2 ± 1.4 mL. Twelve percent of components contained less than 2 mL of RBCs. Mean granulocyte and platelet (PLT) recoveries were 80 and 81%, respectively. There were no transfusion reactions or signs of hemolysis after transfusion of 66 RBC-incompatible granulocyte concentrates (RBC volume, 1.6–8.2 mL). The remaining concentrates were used for topical or intrapleural applications.
RBCs were significantly reduced and granulocytes and PLTs effectively retained in G-CSF/ steroid–mobilized granulocyte components collected with HES and processed by gravity sedimentation. This procedure allows safe transfusion of RBC-incompatible sedimented granulocyte units and may be used to expand the pool of available granulocyte donors for specific recipients.
PMCID: PMC3421031  PMID: 20113453
17.  Optimized Processing of Growth Factor Mobilized Peripheral Blood CD34+ Products by Counterflow Centrifugal Elutriation 
Growth factor-mobilized peripheral blood progenitor cell products were collected from healthy donors and processed by elutriation. Fractions were collected and analyzed for cell count, viability, and blood cell differential. Using modified elutriation procedures, >99% of red blood cells and platelets were removed from apheresis products with high recoveries of total white blood cells and enrichment of CD34+ cells in one fraction. This process is proven to be a feasible method for the initial manipulation associated with primary blood cell therapy products and supports current good manufacturing process and tissue practice-compliant cell processing.
Cell separation by counterflow centrifugal elutriation has been described for the preparation of monocytes for vaccine applications, but its use in other current good manufacturing practice (cGMP) operations has been limited. In this study, growth factor-mobilized peripheral blood progenitor cell products were collected from healthy donors and processed by elutriation using a commercial cell washing device. Fractions were collected for each product as per the manufacturer's instructions or using a modified protocol developed in our laboratory. Each fraction was analyzed for cell count, viability, and blood cell differential. Our data demonstrate that, using standard elutriation procedures, >99% of red blood cells and platelets were removed from apheresis products with high recoveries of total white blood cells and enrichment of CD34+ cells in two of five fractions. With modification of the basic protocol, we were able to collect all of the CD34+ cells in a single fraction. The CD34-enriched fractions were formulated, labeled with a ferromagnetic antibody to CD34, washed using the Elutra device, and transferred directly to a magnetic bead selection device for further purification. CD34+ cell purities from the column were extremely high (98.7 ± 0.9%), and yields were typical for the device (55.7 ± 12.3%). The processes were highly automated and closed from receipt of the apheresis product through formulation of target-enriched cell fractions. Thus, elutriation is a feasible method for the initial manipulations associated with primary blood cell therapy products and supports cGMP and current good tissue practice-compliant cell processing.
PMCID: PMC3659708  PMID: 23197821
Stem cell; CD34+; Immunodeficient mouse; Cellular therapy
18.  From the donor's arm to blood product: a study on bacterial contamination of apheresis platelet concentrates 
Blood Transfusion  2007;5(3):130-133.
Transfusion-associated bacterial infections are a quite frequent collateral effect of administration of platelet concentrates (PC). We carried out a microbiological surveillance of bacterial contamination of apheresis platelet concentrates by studying microbial flora on donors’ arms before and after skin disinfection, through blood cultures with the diversion volume and with the PC.
Materials and methods
Platelet aphereses were carried out using two Haemonetics MCS+ instruments. Cutaneous swabs were examined by the direct plate technique and blood cultures were performed using Bact/ALERT aerobic bottles. In the 5 years from January 2001 to December 2005 we tested 481 PC.
Cutaneous swabs showed significant bacterial growth in 89% of cases before skin disinfection and in 44% after. None of the blood cultures performed on diversion blood was positive, one (0.2%) PC was positive on the fifth day after collection and the presence of a Staphylococcus epidermidis strain was demonstrated.
Our results suggest that the skin disinfection protocol adopted in our structure is not fully satisfactory. The cultures performed on the PC showed a low prevalence of contamination, and the only positive sample was contaminated by a common skin contaminant (S. epidermidis). The culture became positive on the fifth day after collection, but on the second day the PC had been transfused to a patient, without any adverse reaction.
In our experience a culture method using Bact/ALERT aerobic bottles was not able to prevent transfusion of the only contaminated PC identified in this study.
PMCID: PMC2535893  PMID: 19204765
apheresis; platelet concentrates; microbiological contamination; arm disinfection
19.  Annexin V Release and Transmembrane Mitochondrial Potential during Storage of Apheresis-Derived Platelets Treated for Pathogen Reduction 
In vitro function of stored platelet (PLT) con-centrates was analyzed after applying two different techniques of pathogen reduction technology (PRT) treatment, which could increase cellular injury during processing and storage.
Nine triple-dose PLT apheresis donations were split into 27 single units designated to riboflavin-UVB (M) or psoralen-UVA (I) treatment or remained untreated (C). Throughout 8 days of storage, samples were analyzed for annexin V release, the mitochondrial transmembrane potential (Δψ) and some classical markers of PLT quality (pH, LDH release, hypotonic shock response (HSR)).
PLT count and LDH release of all units maintained initial ranges. All units exhibited a decrease in pH and HSR and an increase in annexin V release and Δψ disruption. Notably, throughout the entire storage period, annexin V release re-mained lowest in M units. Throughout 7 days of storage, M units remained comparable to C units (p > 0.05), whereas inferior values were observed with I units. Here, differences to C units reached significance by day 1 (pH: p < 0.0001), day 5 (annexin V release: p < 0.014), and day 7 (HSR, Δψ: p ≤ 0.003). After PRT treatment, annexin V release and Δψ disruption were significantly (p < 0.001) correlated with pH and HSR.
During storage, all units showed a de-crease in HSR and an increase in acidity, annexin V release and Δψ disruption. While M units remained comparable to C units, I units demonstrated significantly inferior values during terminal storage. This could have resulted from differences in PRT treatment or simply be due to differences in storage media and should be analyzed for clinical relevance in future investigations.
PMCID: PMC2914403  PMID: 20737011
Pathogen reduction technology; Platelet in vitro function; Endogenous annexin V; Transmembrane mitochondrial potential; INTERCEPT BLOOD SYSTEM; MIRASOL-PRT
20.  Toward a patient-based paradigm for blood transfusion 
The current “manufacturing paradigm” of transfusion practice has detached transfusion from the clinical environment. As an example, fresh whole blood in large-volume hemorrhage may be superior to whole blood reconstituted from multiple components. Multicomponent apheresis can overcome logistical difficulties in matching patient needs with fresh component availability and can deliver the benefits of fresh whole blood. Because of the different transfusion needs of patients in emerging economies and the vulnerability of these blood systems to emerging infections, fresh whole blood and multicomponent apheresis can better meet patient needs when compared with transplants of the “manufacturing paradigm”. We propose that patient blood management, along with panels of repeat, paid, accredited apheresis and fresh whole-blood donors can be used in emerging economies to support decentralized blood services. This alternative transfusion–medicine paradigm could eventually also be adopted by established economies to focus transfusion medicine on local patient needs and to alleviate the problem of the aging volunteer donor base.
Video abstract
PMCID: PMC3917954  PMID: 24520208
indications; emerging countries; patient blood management
21.  Donation return time at fixed and mobile donation sites 
Transfusion  2011;52(1):127-133.
This study investigated the effect of blood donation environment, fixed or mobile with differing sponsor types, on donation return time.
Data from 2006 through 2009 at six US blood centers participating in the Retrovirus Epidemiology Donor Study-II (REDS-II) were used for analysis. Descriptive statistics stratified by whole blood (WB), plateletpheresis (PP), and double red blood cell (R2) donations were obtained for fixed and mobile locations, including median number of donations and median interdonation interval. A survival analysis estimated median return time at fixed and mobile sites, while controlling for censored return times, demographics, blood center, and mandatory recovery times.
Two-thirds (67.9%) of WB donations were made at mobile sites, 97.4% of PP donations were made at fixed sites, and R2 donations were equally distributed between fixed and mobile locations. For donations at fixed sites only or alternating between fixed and mobile sites, the highest median numbers of donations were nine and eight, respectively, and the shortest model-adjusted median return times (controlling for mandatory eligibility times of 56 and 112 days) were 36 and 30 days for WB and R2 donations, respectively. For PP donations, the shortest model-adjusted median return time was 23 days at a fixed location and the longest was 693 days at community locations.
WB, PP, and R2 donors with the shortest time between donations were associated with fixed locations and those alternating between fixed and mobile locations, even after controlling for differing mandatory recovery times for the different blood donation procedures.
PMCID: PMC3557842  PMID: 21745215
22.  Transient spleen enlargement in peripheral blood progenitor cell donors given G-CSF 
The administration of granulocyte colony-stimulating factor (G-CSF) to peripheral blood progenitor cell (PBPC) donors causes spleen length to increase, but the duration of enlargement is not known. Eighteen healthy subjects were given 10 μg/kg of G-CSF for 5 days and a PBSC concentrate was collected by apheresis. Ultrasound scans were used to assess craniocaudal spleen length before and after G-CSF administration. Mean spleen length increased from a baseline length of 10.7 ± 1.3 cm to 12.1 ± 1.2 cm on the apheresis day (p < 0.001). Ten days after apheresis, spleen length fell to 10.5 ± 1.2 cm and did not differ from baseline levels (p = 0.57), but in 3 subjects remained 0.5 cm greater than baseline length. Increases in spleen length in PBPC donors are transient and reversible.
PMCID: PMC493285  PMID: 15268759
granulocyte colony-stimulating factor; peripheral blood progenitor cells; splenomegaly; spleen
23.  Therapeutic apheresis in peripheral and retinal circulatory disorders 
In microcirculation disorders, the therapeutic apheresis seems to have two different effects. The first, achieved after only a few sessions, is acute, consisting of drastic reduction of blood viscosity and obtained with the use of low-density lipoprotein (LDL) apheresis, rheopheresis, or fibrinogen apheresis. The second effect is long term, or chronic, and needs to be evaluated after a long course of treatment. The mechanisms underlying the chronic effect are still objects of debate and take into account the pleiotropic effects of apheresis. However, it is likely that the acute effect of apheresis mainly influences the functional components of the vascular damage, and so the derived rheological benefit might last only for a short period. The chronic effect, on the contrary, by acting on the morphological alterations of the vascular walls, requires the apheresis treatment to be prolonged for a longer period or even cycles of treatment to be programmed.
PMCID: PMC3374102  PMID: 22528133
LDL apheresis; Rheopheresis; Microcirculation disorders; Peripheral arterial disease
24.  Therapeutic efficacy of different types of platelet concentrates in thrombocytopenic patients 
Platelet Rich Plasma-Platelet concentrate (PRP-PC), Buffy Coat poor-platelet concentrate (BCPC), and Apheresis — PC were prepared and their therapeutic efficacy were assessed in thrombocytopenic patients.
Study design and methods
PRP-PC and BC-PC were prepared from whole blood and Apheresis-PC by automated cell separator. The post transfusion efficacy of transfused platelets was assessed at 1 hour and 20 hours by corrected count increment (CCI) and percentage recovery (PR).
A total of 60 patients’ (20 each for PRP-PC, BC-PC and Apheresis-PC) were enrolled in this study. Forty one patients received therapeutic and nineteen received prophylactic transfusion support. Patients with aplastic anemia 43% (25/60) and acute leukemia 38% (23/60) formed a majority of study population. Platelet dosage of patients’ received PRP-PC, BC-PC and apheresis-PC were 2.4±0.82 × 1011 (mean±SD), 2.2±0.83 × 1011 (mean±SD) and 4.14±1.82 × 1011 (mean±SD) and ranged from 1.16–4.11 × 1011, 1.04−4.20 × 1011 and 1.22−8.90 × 1011 respectively. There was significantly increase in inter-transfusion interval with Apheresis-PC than with PRP-PC and BC-PC recipients [(Mean±S.D.), 4.7±1.33 days Vs 2.7±0.82 days Vs 2.5±0.7 days respectively] (p < 0.05).
Patients transfused with apheresis-PC had received higher platelet dosage than PRP-PC and BC-PC and this difference was statistically significant (p < 0.001). The post transfusion platelet counts and increments at 1 hour and 20 hours were significantly higher with apheresis-PC than PRP-PC and BC-PC (p < 0.001). However, the corrected count increment (CCI) and percentage recovery (PR) in all three groups were comparable. There was significantly increase in inter-transfusion interval with apheresis-PC than PRPPC and BC-PC (p < 0.05).
PMCID: PMC3453166  PMID: 23100935
Random donor platelets; Buffy coat poor-platelet concentrate; Platelet Rich Plasma-Platelet concentrate; Thrombocytopenic patients
25.  Autologous Peripheral Blood-Derived Mononuclear Cells Induced by Erythropoietin Improve Critical Ischemic Limbs 
Annals of Vascular Diseases  2012;5(1):52-60.
Purpose: Efficient and secure collection of CD34+ cells are crucial for the angiogenic therapies. We have developed autologous peripheral blood-mononuclear cell (MNC) transplantation induced by erythropoietin (rhEPO) for critical ischemic limbs.
Methods: Seven patients, including five with arteriosclerosis obliterans, one with Buerger’s disease and one with progressive systemic sclerosis, underwent ten cell therapies. The first administration of rhEPO was performed two weeks before apheresis, and the second administration and blood donation were performed one week before apheresis to activate bone marrow. MNCs including CD34+ cells, isolated from peripheral blood by apheresis, were immediately injected intramuscularly into ischemic limbs.
Results: The number of peripheral blood-CD34 + cells had significantly increased from 1.32 ± 0.83/microL, before the rhEPO induction, to 1.86 ± 0.94/microL, before the apheresis. The number of transplanted MNCs ranged between 0.5 × 109 and 16.5 × 109, and that of CD34+ cells, between 0.1 × 106 and 12.7 × 106, accounting for 0.02%–0.1% of MNCs. There were no serious complications. Finger ulcers with Buerger’s disease were significantly improved one month after the transplantations, but the same or other ulcer(s) appeared 2–6 months later. Three patients had an improvement in rest pain, and one patient extended maximum pain-free walking distance.
Conclusions: Erythropoietin-induced autologous peripheral blood-MNC transplantation is a useful and safe alternative for ischemic limbs.
PMCID: PMC3595914  PMID: 23555486
Keywordserythropoietin; angiogenesis; autologous peripheral blood-derived mononuclear cell transplantation; critical ischemic limbs

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