Background: Within the EU the collection of mononuclear cells (MNC) as starting source for the manufacturing of autologous cell therapies are mainly performed in hospitals or hospital-based outpatient apheresis centers. We report about the challenges to perform the leukapheresis procedure (LA) at a private held medical practice, with specific emphases on safety, cell collection efficiency, and cost-effectiveness.

Background: We explored the effects of repeated plateletpheresis on the platelet P-selectin expression and soluble P-selectin (sP-selectin) concentrations in platelet donors.Methods: Totally 289 platelet donors and 97 first-time whole blood (WB) donors were enrolled from the blood donor registry at the Fujian provincial blood center, China. The accumulative numbers of plateletpheresis in the last 2 y for participants were recorded, and the basal concentrations of platelet count, sP-selectin and total platelet P-selectin (pP-selectin) were determined.Results: Platelet donors had significantly higher basal concentrations of sP-selectin compared to WB donors (24.12 +/- 7.33 ng/mL vs. 20.74 +/- 5.44 ng/mL, P < 0.0001), with no difference in platelet count and pP-selectin concentrations. Increased numbers of platelet donation were correlated with a steady increase of sP-selectin (r = 0.18, P = 0.002). Multivariate regression analysis identified that the frequency of plateletpheresis is an independent factor for the rise of the sP-selectin concentration (t = 2.64, P = 0.009) while no association was found for pP-selectin and platelet count.Conclusions: Repeated plateletpheresis could result in an increased basal concentration of sP-selectin in blood donors whereas not an alteration in the concentrations of total platelet P-selectin. It remains to be determined whether this might be a consequence of endothelial activation or platelet activation or some other phenomenon. (C) 2016 Elsevier B.V. All rights reserved.

BACKGROUND Recent genome-wide association studies in Caucasians suggested that an association exists between the ABO gene locus and soluble levels of P-selectin (sP-selectin). However, it is unclear if the relationship corresponds to the phenotypic expression of ABO groups or is present in different ethnic groups. The aim of this study was to verify this observation at both genotypic and phenotypic levels in a healthy Chinese population.STUDY DESIGN AND METHODS The ABO blood groups were determined by both phenotypes and genotypes in 440 healthy Chinese Han volunteers, while P-selectin levels were evaluated for sP-selectin and total platelet P-selectin (pP-selectin).RESULTS ABO phenotyping and quantitative analysis of individual sP-selectin plasma levels were combined to demonstrate that individuals phenotypically expressing the A antigen have approximately 20% lower sP-selectin plasma levels than those carrying the B or O phenotype (p<0.0001), but that no difference exists between A and AB and between B and O phenotypes. Genotyping data revealed that the presence of the A gene could be attributed to the observed difference in phenotype comparison, with no difference between A/A, A/B, and A/O genotypes. There were also no associations between ABO blood groups, either phenotypes or genotypes, and pP-selectin levels.CONCLUSION This study demonstrated an association between sP-selectin levels and ABO groups in a Chinese Han population, implicating its generalizability to other ethnic groups. This finding will improve the understanding of the mechanism of ABO blood group-associated diseases.

This study aims to assess the association of the preoperative neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) with tumor stage in colorectal cancer (CRC) patients. A retrospective study was performed in 336 CRC patients. Preoperative whole blood counts, serum levels of carcinoembryonic antigen (CEA), and clinicopathologic data were collected. The correlations between laboratory parameters and the tumor, node, and metastasis (TNM) stages were analyzed. The clinicopathologic TNM stages among CRC patients were 12.8 % at stage I, 32.4 % at stage II, 44.6 % at stage III, and 10.1 % at stage IV. NLR, PLR, and CEA levels were higher in CRC patients compared to healthy controls (all P < 0.0001). Both NLR and PLR showed an early elevation as compared to CEA, with a higher area under curve (AUC) value (0.71 vs. 0.62) in predicting the presence of the tumor with stage I/II. Accordingly, significant elevations of NLR (P = 0.0018) and PLR (P < 0.0001) were firstly detected in stage I and stage II, respectively. In addition, NLR exhibited a second phase elevation in stage IV, with a significant higher level in M1 subgroup compared to M0 subgroup (P = 0.022). While PLR showed a T stage-dependent increase (P = 0.0003) and was identified as an independent factor for the T grade development (P < 0.0001). Our data indicated that both neutrophil- and platelet-mediated inflammatory reactions are predominantly involved in the different stages of CRC development. Determination of pretreatment levels of NLR and PLR might provide useful information for the early diagnosis or the therapeutic choices in CRC patients.

Mononuclear cell (MNC) preparations collected by leukapheresis are used as cellular source for the ex vivo generation of immunotherapeutic approaches. Propose of the present analysis was to compare MNC kinetics during non-stimulated leukapheresis in patients with advanced cancer, with particular focus on the influence of chemotherapy on changes in peripheral blood cell counts, cell recruitment, and the quality of the collected cell preparations.

BACKGROUND AIMSLittle is known of the effect of anticoagulation on peripheral blood progenitor cell (PBPC) harvest during large-volume leukapheresis (LVL). Because of the interaction of heparin with stromal cell-derived factor (SDF)-1α, it has been proposed that a heparin-based anticoagulation may result in an increased PBPC collection efficiency compared with standard citrate-based anticoagulation.METHODSWe conducted a prospective randomized trial to address the effect of both anticoagulation regimes on safety, subjective comfort and CD34 (+) collection efficiency in 90 adult patients undergoing standardized LVL. Anticoagulation consisted of either citrate (group C) or a combination of heparin and low-dose citrate (group H).RESULTSThe overall incidence of adverse reactions (AR) during LVL was 17%. AR consisted only of citrate-related AR; no bleeding complications were observed. Determination of parameters of the acid-base balance revealed a higher frequency of metabolic alkalosis in group C. Analysis of serum SDF-1α revealed no differences in SDF-1α plasma levels. There were no differences in the CD34 (+) cell collection efficiency, resulting in the harvest of equal CD34 (+) cell yields independent of the anticoagulation used.CONCLUSIONSOur data show no clinical relevant effect of a heparin containing anticoagulation in terms of an increased overall CD34 (+) cell collection during LVL, although this regime shows some benefits in terms of the incidence and subjective tolerance towards AR. Based on our results the decision between a citrate- and heparin-substituted anticoagulation for LVL should be driven by patient-related factors, and should concern potential contraindications of both methods.

BACKGROUND AIMS:Bone marrow (BM)-derived mononuclear cell (MNC) preparations are increasingly used in experimental studies exploring the potential effect of progenitor cell-derived therapies in cardiocirculatory diseases. We analyzed the cellular BM composition, side-effects and other process-related variables of BM harvest and BM-MNC preparation in 80 patients with cardiovascular disease.METHODS:BM (median 828 mL, range 223-1038 mL) was collected from the iliac crest. After BM harvest the MNC fraction was enriched by semi-automatic apheresis to reduce the total volume of the transplant. Autologous red blood cells (RBC) were salvaged from the initial BM harvest and autotransfused to the patients.RESULTS:There were no serious side-effects related to BM collection, particularly no serious bleeding complications. Twenty- five of 80 (31%) patients developed mild pain. BM harvest resulted in the collection of a median of 2.8 × 10(9) MNC, containing a median of 66.5 × 10(6) CD34/45 cells, 39.5 × 10(6) CD133/45 cells and 50.3 × 10(6) CD34/CD133 cells. Apheresis technology-based MNC enrichment of harvested BM resulted in a progenitor cell recovery of 69-75.3% of total cells. Additional salvage of RBC from the initial BM harvest resulted in the recovery of a median of 175.0 mL autologous RBC mass. Transfusion of salvaged RBC was well tolerated and resulted in a significant increase in hemoglobin levels.CONCLUSIONS:Collection of BM of up to 1 L in combination with in vitro processing using a semi-automated apheresis device is a safe and feasible approach to increasing the number of progenitor cells necessary for cellular therapies, particularly when combined with RBC salvage.

PURPOSE:Several observational studies in head and neck cancer have reported that allogenic blood transfusion is associated with increased postoperative complications, increased risk of tumor recurrence, and worse prognosis. The aim of this study was to identify preoperative and intraoperative factors predicting blood transfusion in patients undergoing surgery for oral and oropharyngeal cancer. PATIENTS AND METHODS:We conducted a retrospective cohort study of patients undergoing tumor resection and free flap reconstruction for locally advanced oral and oropharyngeal squamous cell carcinoma between 2000 and 2008. The primary outcome variable was perioperative exposure to allogenic blood transfusion. Univariate and multivariate logistic regression models were used to determine predictors of blood transfusion. RESULTS:A cohort of 142 participants was found eligible. In a multivariate model, Charlson score ≥ 1 (OR, 5.2; 95% CI, 1.4 to 19.3; P = .01), preoperative hemoglobin levels ≤ 12 g/dl (OR, 4.4; 95% CI, 1.2 to 16.2; P = .03), bone resection (OR, 5.1; 95% CI, 1.5 to 17.8; P = .01), and osseous free tissue transfer (OR, 8.8; 95% CI, 1.0 to 74.8; P = .046) were independently associated with an increased risk of blood transfusion. CONCLUSION:Our study identified patient- and surgery-related factors predicting a higher risk of exposure to allogenic blood transfusion. This readily available preoperative information could be used to better stratify patients according to their transfusion risk and may thereby guide blood conservation strategies in high-risk patients.

Background: Citrate is the anticoagulation of choice in apheresis procedures. Citrate anticoagulation results in a short-term increase in serological markers of bone turnover, with uncertain clinical significance.Aim: To understand the effect of calcium supplementation on serological bone turnover markers during an acute citrate load as a mimic of citrate anticoagulation during apheresis procedures.Methods: A placebo-controlled, crossover study was conducted in 22 healthy volunteers. Volunteers received a standardized citrate load at a fixed dose of 1.5 mg/kg of body weight/min for 80 min for three times and a single placebo infusion as a control. Each intervention was separated by a wash-out interval of 2 to 3 weeks. During two citrate infusions, volunteers received an additional calcium supplementation, consisting of either oral administration of calcium carbonate or an i.v. bypass infusion of calcium gluconate. Serial blood samples were collected for the determination of ionized calcium (iCa), intact parathyroid hormone (iPTH) and markers of bone remodeling, C-telopeptide of type I collagen (CTX) and osteocalcin (OC).Results: The infusion of citrate without calcium supplementation resulted in an increase in the bone formation marker OC and the bone resorption marker CTX, in addition to the changes in iPTH and iCa. The administration of calcium by either oral administration or as an i.v, bypass infusion attenuated the observed changes in CTX, but showed no effects on the elevation of the bone formation marker OC. There was no difference in the attenuation of CTX between the two calcium formulations. However, the i.v. application of calcium gluconate had a superior effect in reducing the change of serum iPTH and iCa as compared to the oral administration of calcium carbonate.Conclusions: Calcium supplementation is an effective method in damping the citrate-related transient increase of the serological bone resorption marker CTX. As a mimic for the citrate-based apheresis procedure, our data may enforce the prophylactic application of calcium supplementation to attenuate the short-term elevation of bone resorption related to an acute citrate load. (C) 2011 Elsevier Inc. All rights reserved.

Evidence indicates that allogenic packed red blood cell transfusion results in the host’s immunomodulation, and is associated with adverse clinical outcomes after surgery. The aim of this study was to test whether allogenic leukocyte-depleted blood transfusion represents a significant risk factor for postoperative morbidity after oral and oropharyngeal cancer surgery. A total of 142 patients, diagnosed for the first time with oral and oropharyngeal squamous cell carcinoma, and receiving neoadjuvant chemoradiotherapy followed by surgery between 2000 and 2008 were retrospectively included in this study. Univariate and multivariate logistic regression models were calculated to identify predictors of postoperative complications. We found a significantly higher complication rate in the group of transfused patients compared to patients not exposed to transfusion (complication rate of 84% and 39%, respectively, p < 0.001). On multivariate analysis, the amount of packed red blood cells transfused (for 1–4 units transfused: adjusted OR, 2.59; 95% CI, 1.24–5.39; p = 0.011; for more than >4 units transfused: adjusted OR, 5.29; 95% CI, 2.01–13.88; p = 0.001) and Charlson’s comorbidity score ⩾1 (adjusted OR, 2.81; 95% CI, 1.38–5.70; p < 0.004) were independently associated with the development of postoperative complications. Allogenic leukocyte-depleted blood transfusion is independently associated with increased postoperative complications in patients undergoing surgery for oral and oropharyngeal cancer. This association follows a dose–response relationship, as patients who received larger amounts of packed red blood cells showed a significant trend toward higher postoperative morbidity.

BackgroundAfter large volume bone marrow (BM) harvest, donors and patients can develop severe anaemia, because collected BM can contain up to 20% of their red cell mass. In a prospective analysis, we investigated the feasibility to recover red blood cells (RBCs) from the harvested BM and investigated whether these RBC units meet the quality requirements of the European Council.Patients and MethodsFrom 19 patients (median age 51 yrs, range 31-77) with acute myocardial infarction, who participated in the MYSTAR study, a median volume of 1299 ml (range, 700-1870 ml) BM was collected. During BM processing, mononuclear cells (MNC) were separated using the Cobe Spectra (TM) apheresis system and the residual RBCs were collected in a separate bag. The quality of the collected RBCs was assessed by measuring LDH, free haemoglobin, potassium and lactate. Haemolysis was calculated and the intracellular concentration of ATP, ADP, AMP was determined by HPLC.ResultsRBC units recovered from BM after MNC separation had a mean volume of 312 +/- 95 ml with a haematocrit of 47 +/- 8 center dot 9%, a haemoglobin content of 51 +/- 15 g per unit, a haemolysis of 0 center dot 15 +/- 0 center dot 005%, a pH of 6 center dot 8 +/- 0 center dot 007 and an intracellular ATP concentration of 135 pmol/106 RBC +/- 41, which is comparable with freshly collected packed red blood cells (PRBCs).ConclusionRBCs, collected from bone marrow harvests, can be used for autologous blood support to minimize allogeneic blood transfusions in donors and patients after large volume BM donation.

SummaryThe aim of the sub-study of the MYSTAR randomised trial was to analyse the changes in myocardial perfusion in NOGA-defined regions of interest (ROI) with intramyocardial injections of autologous bone marrow mononuclear cells (BM-MNC) using an elaborated transformation algorithm. Patients with recent first acute myocardial infarction (AMI) and left ventricular (LV) ejection fraction (EF) between 30–45% received BM-MNC by intramyocardial followed by intracoronary injection 68 ± 34 days post-AMI (pooled data of MYSTAR). NOGA-guided endocardial mapping and 99m-Sestamibi-SPECT (single photon emission computer tomography) were performed at baseline and at three months follow-up (FUP). ROI was delineated as a best polygon by connecting of injection points of NOGA polar maps. ROIs were projected onto baseline and FUP polar maps of SPECT calculating the perfusion severity of ROI. Infarct size was decreased (from 27.2 ± 10.7% to 24.1 ± 11.5%, p<0.001), and global EF increased (from 38 ± 6.1% to 41.5 ± 8.4%, p<0.001) three months after BM-MNC delivery. Analysis of ROI resulted in a significant increase in unipolar voltage (index of myocardial viability) (from 7.9 ± 3.0 mV to 9.9 ± 2.7 mV at FUP, p<0.001) and local linear shortening (index of local wall motion disturbances) (from 11.0 ± 3.9% to 12.7 ± 3.4%, p=0.01). NOGA-guided analysis of the intramyocardially treated area revealed a significantly increased tracer up-take both at rest (from 56.7 ± 16.1% to 62.9 ± 14.2%, p=0.003) and at stress (from 59.3 ± 14.2% to 62.3 ± 14.9%, p=0.01). Patients exhibiting ≥5% improvement in perfusion defect severity received a significantly higher number of intramyocardial BM-MNC. In conclusion, combined cardiac BM-MNC delivery induces significant improvement in myocardial viability and perfusion in the intramyocardially injected area.Clinical Trials Gov Nr: NCT00384982.

Objective To investigate the possible effect of citrate on electrolyte metabolism in healthy people with different genders and races and provide a reference for the possible clinical interventions.Methods A cross over,placebo-controlled study was conducted in 22 age-matched Chinese(11 males and 11 females)and 10 male Caucasian volunteers after informed consents were obtained.Volunteers received of saline solution,separated by a wash-out period of two to three weeks.Serial blood and urine samples were collected during the observation period and analyzed for the selective biochemical parameters.Results Comparable basal levels of serum albumin[male(43.05±1.81)g/L vs female(42.26±2.67)g/L]and serum ionized calcium[male(1.27±0.04)mmol/L vs female(1.26±0.04)mmol/L]were observed between different genders of Chinese volunteers.However,citrate intervention led to more pronounced decrease of ionized calcium level in Chinese females compared to Chinese males[-28.68%(-20.00%--35.2%)vs-23.84%(-16.53%--29.32%),t=3.19,P ＜ 0.01].There was no differences of the levels of serum inorganic phosphate[-18.81%(-3.16%--25.09%)vs-19.23%(-1.22%--32.16%),t=0.36,P＞0.05]and albumin[-0.32%(3.27%--7.60%)vs 1.88%(6.03%--9.31%),t=0.47,P＞0.05].Independent of gender,citrate intervention resulted in an increased excretion of urine calcium in Chinese volunteers[before 0.34(0.09-0.87)vs after 0.96(0.18-1.47),t=6.66,P ＜0.01].Compared to Caucasian males,Chinese males has a higher basal level of serum ionized calcium [(1.27±0.04)mmol/L vs(1.22±0.02)mmol/L,t=3.7,P ＜0.01]and larger amplitude basal rhythm in serum albumin level[-11.72%(-5.70%--14.21%)vs-1.74%(2.43%--7.68%),t=7.43,P ＜ 0.01].Application of citrate resulted in comparable changes of serum ionized calcium [-23.84%(-16.53%--29.32%)vs-21.95%(-18.31%--30.92%)],phosphate[-19.23%(4.65%--32.16%)vs-12.68%(0.68%--42.19%)],albumin[-0.32%(1.05%--7.60%)vs-1.39%(1.87%--7.26%)]and urine calcium excretion[237.70%(11.8%-935%)vs 234.37%(5.45%-504.00%)]between Chinese and Caucasian males(t=0.32,0.03,0.25 and 0.04 respectively,P＞0.05).Serum levels of magnesium were not influenced in all volunteers during two interventions.Conclusions Independent of race and gender,the invention of citrate results in comparable changes of serum magnesium,inorganic phosphate and albumin.The effect of citrate on ionized calcium levels between genders implicates a higher risk for hypocalcemic reactions in females compared to males undergoing automatic apheresis procedures.

目的 了解利用血细胞分离机大处理量地富集外周血单核细胞技术在临床肿瘤患者免疫治疗应用中的效果和可行性.方法 利用血细胞分离机对患者进行大处理血量外周血单个核细胞(MNC)采集以富集单核细胞;并同时对患者采集前后外周全血细胞计数、采集产物的组成及采集相关不良反应进行分析.结果 采集全过程处理血量为患者全身血容量的3.17倍,MNC 产物中单核细胞含量达30.3%,绝对细胞数达2.80×109个,采集效率为50.5%.采集产物中单核细胞的产量与患者采集前外周血白细胞的计数高低相关.该采集过程导致的不良反应是一定程度的白细胞和血小板的丢失.采集后患者外周血白细胞和血小板计数分别下降12.47%和19.27%,但无明显的抗凝剂相关不良反应发生.结论 应用血细胞分离机大处理量地富集外周血单核细胞可获得足以满足DC治疗需要的临床级单核细胞产量.

Vox SanguinisVolume 97, Issue 1 p. 77-90 New cellular therapies: Is there a role for transfusion services? H. W. Reesink, H. W. ReesinkSearch for more papers by this authorS. Panzer, S. PanzerSearch for more papers by this authorM. Dettke, M. DettkeSearch for more papers by this authorC. Gabriel, C. GabrielSearch for more papers by this authorM. Lambermont, M. LambermontSearch for more papers by this authorV. Deneys, V. DeneysSearch for more papers by this authorD. Sondag, D. SondagSearch for more papers by this authorE. Dickmeiss, E. DickmeissSearch for more papers by this authorA. Fischer-Nielsen, A. Fischer-NielsenSearch for more papers by this authorM. Korhonen, M. KorhonenSearch for more papers by this authorT. Krusius, T. KrusiusSearch for more papers by this authorA. Ali, A. AliSearch for more papers by this authorP. Tiberghien, P. TiberghienSearch for more papers by this authorH. Schrezenmeier, H. SchrezenmeierSearch for more papers by this authorT. Tonn, T. TonnSearch for more papers by this authorE. Seifried, E. SeifriedSearch for more papers by this authorH. Klüter, H. KlüterSearch for more papers by this authorC. Politis, C. PolitisSearch for more papers by this authorA. Stavropoulou-Gioka, A. Stavropoulou-GiokaSearch for more papers by this authorM. Parara, M. PararaSearch for more papers by this authorØ. Flesland, Ø. FleslandSearch for more papers by this authorF. Nascimento, F. NascimentoSearch for more papers by this authorB. Balint, B. BalintSearch for more papers by this authorP. Marin, P. MarinSearch for more papers by this authorT. Bart, T. BartSearch for more papers by this authorF. E. Chen, F. E. ChenSearch for more papers by this authorD. H. Pamphilon, D. H. PamphilonSearch for more papers by this author H. W. Reesink, H. W. ReesinkSearch for more papers by this authorS. Panzer, S. PanzerSearch for more papers by this authorM. Dettke, M. DettkeSearch for more papers by this authorC. Gabriel, C. GabrielSearch for more papers by this authorM. Lambermont, M. LambermontSearch for more papers by this authorV. Deneys, V. DeneysSearch for more papers by this authorD. Sondag, D. SondagSearch for more papers by this authorE. Dickmeiss, E. DickmeissSearch for more papers by this authorA. Fischer-Nielsen, A. Fischer-NielsenSearch for more papers by this authorM. Korhonen, M. KorhonenSearch for more papers by this authorT. Krusius, T. KrusiusSearch for more papers by this authorA. Ali, A. AliSearch for more papers by this authorP. Tiberghien, P. TiberghienSearch for more papers by this authorH. Schrezenmeier, H. SchrezenmeierSearch for more papers by this authorT. Tonn, T. TonnSearch for more papers by this authorE. Seifried, E. SeifriedSearch for more papers by this authorH. Klüter, H. KlüterSearch for more papers by this authorC. Politis, C. PolitisSearch for more papers by this authorA. Stavropoulou-Gioka, A. Stavropoulou-GiokaSearch for more papers by this authorM. Parara, M. PararaSearch for more papers by this authorØ. Flesland, Ø. FleslandSearch for more papers by this authorF. Nascimento, F. NascimentoSearch for more papers by this authorB. Balint, B. BalintSearch for more papers by this authorP. Marin, P. MarinSearch for more papers by this authorT. Bart, T. BartSearch for more papers by this authorF. E. Chen, F. E. ChenSearch for more papers by this authorD. H. Pamphilon, D. H. PamphilonSearch for more papers by this author First published: 07 June 2009 https://doi.org/10.1111/j.1423-0410.2009.01184.xCitations: 5AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume97, Issue1July 2009Pages 77-90 RelatedInformation