RESUMO
BACKGROUND: High-dose chemotherapy followed by autologous hematopoietic stem cell transplantation (ASCT) is routinely used in various hematologic malignancies. However, dimethylsulfoxide contained in cryopreserved grafts can cause adverse events (AEs). STUDY DESIGN AND METHODS: Forty-three ASCTs were performed with Sepax 2 washed grafts between 7/2016 and 10/2019. The aim of this study was to determine whether washing out dimethyl sulfoxide (DMSO) from transplants using the Sepax 2 (S-100) device is safe and reduces the incidence of DMSO-associated AEs. RESULTS: The washing procedure was automated and that resulted in the satisfactory recovery of total nucleated cells, CD34+ cells, and colony forming units of granulocyte and macrophages (85%, 80%, and 84%, medians). Time to engraftment of leukocytes, granulocytes, and platelets as well as the number of neutropenic days did not differ when compared to 20 consecutive ASCTs without washing. The AE occurrence was lower compared to unwashed grafts: 81% versus 78% during and shortly after grafts administration, 76% versus 69% in the following day. CONCLUSION: We conclude that the washing of cryopreserved transplants using Sepax 2 was feasible with a high recovery of hematopoietic cells, did not influence time to engraftment, and resulted in the satisfactory reduction of AEs and improved tolerance of the procedure.
Assuntos
Crioprotetores/efeitos adversos , Dimetil Sulfóxido/efeitos adversos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Adulto , Idoso , Criopreservação/instrumentação , Criopreservação/métodos , Crioprotetores/isolamento & purificação , Dimetil Sulfóxido/isolamento & purificação , Feminino , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/citologia , Humanos , Masculino , Pessoa de Meia-Idade , Transplante AutólogoRESUMO
BACKGROUND: CD34+ cell count correlates with engraftment potency after autologous stem cell transplantation. Assessment of CD34+ mainly occurs after apheresis and before cryopreservation with dimethyl sulfoxide (DMSO). The influence of postthaw CD34+ cell numbers over time to engraftment is not well studied, and determination of postthaw CD34+ cell counts is challenging for a variety of reasons. The aim of this retrospective study was to systematically assess the value of postthaw CD34+ cell counts in autologous grafts with and without DMSO removal. STUDY DESIGN AND METHODS: Between January 2008 and December 2015, 236 adult patients underwent a total of 292 autologous stem cell transplantations. Median age at transplantation was 56 years, and the main indication was multiple myeloma (60%). DMSO removal was done in 96 grafts (33%), either by centrifugation or by Sepax method. RESULTS: Patients receiving grafts containing DMSO showed a significantly faster platelet (p = 0.02) and RBC (p = 0.001) engraftment. DMSO removal was not associated with fewer infusion-related adverse events. We observed a good correlation between CD34+ cell count after apheresis and CD34+ cell count after thawing/washing (r = 0.931). Ninety grafts (31%) showed a significant loss of viable CD34+ cells, which translated into a delayed engraftment. CONCLUSION: DMSO removal was associated with delayed platelet and RBC engraftment without preventing adverse events. CD34+ cell enumeration after thawing remains difficult to perform, but grafts showing higher cell loss during cryopreservation and thawing are associated with slower engraftment. Prospective studies on the role of DMSO removal and postthaw CD34+ enumeration using defined protocols are needed.
Assuntos
Antígenos CD34/análise , Dimetil Sulfóxido/isolamento & purificação , Transplante de Células-Tronco Hematopoéticas/métodos , Transplante Autólogo/métodos , Adolescente , Adulto , Idoso , Criopreservação , Crioprotetores/isolamento & purificação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto JovemRESUMO
BACKGROUND: Cryopreserved hematopoietic progenitor cell (HPC) grafts are widely infused to patients with malignant and nonmalignant conditions. Despite reduction of immediate side effects linked to dimethyl sulfoxide (DMSO), cell debris-containing grafts and comparable hematopoietic engraftment between washed and unwashed cryopreserved products, bedside infusion of thawed HPC grafts is still preferred. Introduction of automated devices is important for standardization and consistency of graft manipulation. Additionally, these techniques are likely to be useful for the delivery of innovative cell-based medicinal products that are currently under development. METHODS: In this study, we evaluated three consecutive versions of the Lovo device (Fresenius Kabi) for automated washing of thawed HPC products. A total of 42 HPC products intended for destruction were used. Measured outcomes included viable CD34+ cell recovery, viability, total processing time and post-washing stability. RESULTS: Preliminary data using the prototype Lovo 0.0 to process a single HPC unit showed better recovery and viability of CD34+ cells using a two-cycle than a three-cycle wash, with >95% DMSO elimination. The Lovo 1.0 performed equally well. When simultaneously processing two HPC units, the upgraded Lovo 2.0 device demonstrated comparable CD34+ recovery, DMSO elimination efficiencies and time-saving capacity. Furthermore, washed cell products were stable for 4 hours at room temperature. DISCUSSION: Lovo device satisfies clinically relevant issues: ability to efficiently wash two HPC units simultaneously and compatibility with transport to nearby transplantation centers.
Assuntos
Criopreservação/instrumentação , Dimetil Sulfóxido/isolamento & purificação , Células-Tronco Hematopoéticas/citologia , Antígenos CD34/metabolismo , Criopreservação/métodos , Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/metabolismo , Humanos , TemperaturaRESUMO
BACKGROUND: Preparation of umbilical cord blood units (CBUs) for infusion requires a step of dilution or washing to reduce the toxicity of the dimethyl sulfoxide present in the freezing solution. However, the worldwide shortage of clinical-grade dextran 40, the most widely used cord blood dilution or washing solution, prompted us to search for an alternative solution. We elected to evaluate the performance of alternative solutions that could be used as potential replacements for the dextran 40-based solution. STUDY DESIGN AND METHODS: Frozen CBUs were rapidly thawed and immediately diluted 1:4 in each of 10 dilution solution variants of dextran 40, Plasma-lyte, or Hespan. We compared these alternatives by assessing viability, CD34, CD45, and colony-forming units recovery postthaw. RESULTS: A significantly lower CD34 and CD45 recovery was observed in all solutions without human serum albumin (HSA). All solutions with 5% HSA gave recovery, viability, and potency figures similar to the dextran 40/HSA solution. CONCLUSION: Based on our results and considering that Plasma-lyte A (PA)/HSA is already used for the thawing of CD34 from mobilized blood, we conclude that PA/HSA could be a safe and efficient solution for the replacement of dextran 40-based dilution solutions.
Assuntos
Criopreservação , Dextranos/farmacologia , Sangue Fetal/efeitos dos fármacos , Substitutos do Plasma/farmacologia , Antígenos CD34/análise , Sobrevivência Celular , Dimetil Sulfóxido/isolamento & purificação , Eletrólitos/farmacologia , Humanos , Derivados de Hidroxietil Amido/farmacologia , Antígenos Comuns de Leucócito/análise , Células-TroncoRESUMO
Transplantation of hematopoietic stem cells (HSCs) has been successfully developed as a part of treatment protocols for a large number of clinical indications, and cryopreservation of both autologous and allogeneic sources of HSC grafts is increasingly being used to facilitate logistical challenges in coordinating the collection, processing, preparation, quality control testing and release of the final HSC product with delivery to the patient. Direct infusion of cryopreserved cell products into patients has been associated with the development of adverse reactions, ranging from relatively mild symptoms to much more serious, life-threatening complications, including allergic/gastrointestinal/cardiovascular/neurological complications, renal/hepatic dysfunctions, and so on. In many cases, the cryoprotective agent (CPA) used-which is typically dimethyl sulfoxide (DMSO)-is believed to be the main causal agent of these adverse reactions and thus many studies recommend depletion of DMSO before cell infusion. In this paper, we will briefly review the history of HSC cryopreservation, the side effects reported after transplantation, along with advances in strategies for reducing the adverse reactions, including methods and devices for removal of DMSO. Strategies to minimize adverse effects include medication before and after transplantation, optimizing the infusion procedure, reducing the DMSO concentration or using alternative CPAs for cryopreservation and removing DMSO before infusion. For DMSO removal, besides the traditional and widely applied method of centrifugation, new approaches have been explored in the past decade, such as filtration by spinning membrane, stepwise dilution-centrifugation using rotating syringe, diffusion-based DMSO extraction in microfluidic channels, dialysis and dilution-filtration through hollow-fiber dialyzers and some instruments (CytoMate, Sepax S-100, Cobe 2991, microfluidic channels, dilution-filtration system, etc.) as well. However, challenges still remain: development of the optimal (fast, safe, simple, automated, controllable, effective and low cost) methods and devices for CPA removal with minimum cell loss and damage remains an unfilled need.
Assuntos
Criopreservação/métodos , Crioprotetores/efeitos adversos , Dimetil Sulfóxido/efeitos adversos , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Transplante de Células-Tronco Hematopoéticas/métodos , Crioprotetores/isolamento & purificação , Dimetil Sulfóxido/isolamento & purificação , Humanos , Condicionamento Pré-Transplante/efeitos adversos , Transplante HomólogoRESUMO
BACKGROUND: Products cryopreserved with dimethyl sulfoxide (DMSO) in stem cell transplant (SCT) often cause many adverse effects during their infusion (major cardiovascular events, dyspnea even death). These are especially frequent in pediatric patients. We tested if a fully automated and closed wash procedure (Sepax S-100, Biosafe) allowed us to maintain the absolute CD34+ cell number, cell viability, and engraftment potential, decreasing the untoward reactions. STUDY DESIGN AND METHODS: Forty-six washes of DMSO cryopreserved peripheral blood hematopoietic progenitor (HP) apheresis were studied. Blood aliquots were taken both after thawing and after washing to assess the total nucleated and CD34+ cell counts, as well as cell viability. The washed products were infused in 26 autologous SCTs (ASCTs). Results were compared with the 53 previous SCTs performed without DMSO removal. RESULTS: After washing there were no significant differences between the pre- and postwashing CD34+ cell counts (p=0.08) or viability (p=0.68). No significant differences were observed between washed and nonwashed infusions in relation to the day of the neutrophil (p=0.46) and platelet (p=0.26) engraftment. One adverse event, abdominal pain, occurred during the washed cells infusions. When compared with the 14 untoward reactions that took place during the nonwashed HP infusions, significance was reached (p=0.00043). CONCLUSIONS: The automatic method described is effective in terms of CD34+ cell recovery and viability in ASCT. Moreover, Sepax decreased significantly the untoward reactions during the infusion.
Assuntos
Preservação de Sangue/efeitos adversos , Crioprotetores/isolamento & purificação , Dimetil Sulfóxido/isolamento & purificação , Transplante de Células-Tronco Hematopoéticas/normas , Doença de Hodgkin/terapia , Adulto , Idoso , Remoção de Componentes Sanguíneos , Preservação de Sangue/métodos , Transfusão de Sangue Autóloga , Sobrevivência Celular/efeitos dos fármacos , Criança , Pré-Escolar , Crioprotetores/efeitos adversos , Dimetil Sulfóxido/efeitos adversos , Feminino , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Humanos , Linfoma não Hodgkin/terapia , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/terapiaRESUMO
Cells are routinely cryopreserved for investigative and therapeutic applications. The most common cryoprotective agent (CPA), dimethyl sulfoxide (DMSO), is toxic, and must be removed before cells can be used. This study uses a microfluidic device in which three streams flow vertically in parallel through a rectangular channel 500 µm in depth. Two wash streams flow on either side of a DMSO-laden cell stream, allowing DMSO to diffuse into the wash and be removed, and the washed sample to be collected. The ability of the device to extract DMSO from a cell stream was investigated for sample flow rates from 0.5 to 4.0 mL/min (Pe = 1,263-10,100). Recovery of cells from the device was investigated using Jurkat cells (lymphoblasts) in suspensions ranging from 0.5% to 15% cells by volume. Cell recovery was >95% for all conditions investigated, while DMSO removal comparable to a previously developed two-stream device was achieved in either one-quarter the device length, or at four times the flow rate. The high cell recovery is a ~25% improvement over standard cell washing techniques, and high flow rates achieved are uncommon among microfluidic devices, allowing for processing of clinically relevant cell populations.
Assuntos
Criopreservação/métodos , Crioprotetores/isolamento & purificação , Dimetil Sulfóxido/isolamento & purificação , Técnicas Analíticas Microfluídicas/instrumentação , Movimento Celular/fisiologia , Tamanho Celular , Criopreservação/instrumentação , Crioprotetores/química , Difusão , Dimetil Sulfóxido/química , Desenho de Equipamento , Humanos , Células Jurkat , Técnicas Analíticas Microfluídicas/métodos , Modelos BiológicosRESUMO
Fenton catalytic degradation of dimethyl sulfoxide (DMSO) by using liquid-phase pulsed high voltage discharge was carried out. The discharge was driven by a self-made pulsed power supply providing a pulse rising time of 400 ns, discharge repetition rate of 96.2 Hz, peak voltage of 20 kV. The side surface of discharge electrode was insulated in order to limit the discharge current, followed by estimating its effect on single pulse power. The effects of aqueous conductivity, Fe(II), and O2 flow rate on liquid-phase discharge induced DMSO degradation and aqueous conductivity on corresponding H2O2 yield were investigated. The concentrations and selectivities of intermediates of DMSO degradation, were also studied. The results indicated that when using the insulated electrode, single pulse power had a limiting value with increasing the aqueous conductivity; DMSO degradation rate was reduced with increasing the aqueous conductivity and O2 flow rate,whereas Fe(II) showed a Fenton catalytic oxidation of DMSO; 80% of DMSO degradation rate was achieved at the pulsed high voltage discharge time of 45 min and the resultant biodegradability was enhanced by at least 32%-48%; the maximum energy efficiency in G(37%) was 0.0087 mol/(kW x h). The study suggested that the liquid-phase plasma combined catalyst promised the potential of organic compound degradation.
Assuntos
Dimetil Sulfóxido/química , Dimetil Sulfóxido/isolamento & purificação , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da Água/isolamento & purificação , Catálise , Eletroquímica/métodos , Eletrodos , Oxirredução , Poluentes Químicos da Água/químicaRESUMO
The mild catalytic oxidation of dimethylsulfoxide (DMSO) into biodegradable dimethylsulfone is proposed as an efficient pretreatment of wastewaters subjected to biological treatment processes. A SiO(2)-TiO(2) mesoporous xerogel prepared by a non-hydrolytic route, as well as titanium silicalite TS-1 showed very good activity and stability in the catalytic oxidation of DMSO with H(2)O(2) in dilute aqueous solution, at room temperature.
Assuntos
Dimetil Sulfóxido/química , Dimetil Sulfóxido/isolamento & purificação , Peróxido de Hidrogênio/química , Titânio/química , Eliminação de Resíduos Líquidos , Poluentes Químicos da Água/química , Poluentes Químicos da Água/isolamento & purificação , Catálise , Oxirredução , Dióxido de Silício/química , TemperaturaRESUMO
The amount of pollutants produced during manufacturing processes of TFT-LCD (thin-film transistor liquid crystal display) substantially increases due to an increasing production of the opto-electronic industry in Taiwan. The total amount of wastewater from TFT-LCD manufacturing plants is expected to exceed 200,000 CMD in the near future. Typically, organic solvents used in TFT-LCD manufacturing processes account for more than 33% of the total TFT-LCD wastewater. The main components of these organic solvents are composed of the stripper (dimethyl sulphoxide (DMSO) and monoethanolamine (MEA)), developer (tetra-methyl ammonium hydroxide (TMAH)) and chelating agents. These compounds are recognized as non-or slow-biodegradable organic compounds and little information is available regarding their biological treatability. In this study, the performance of an A/O SBR (anoxic/oxic sequencing batch reactor) treating synthetic TFT-LCD wastewater was evaluated. The long-term experimental results indicated that the A/O SBR was able to achieve stable and satisfactory removal performance for DMSO, MEA and TMAH at influent concentrations of 430, 800, and 190 mg/L, respectively. The removal efficiencies for all three compounds examined were more than 99%. In addition, batch tests were conducted to study the degradation kinetics of DMSO, MEA, and TMAH under aerobic, anoxic, and anaerobic conditions, respectively. The organic substrate of batch tests conducted included 400 mg/L of DMSO, 250 mg/L of MEA, and 120 mg/L of TMAH. For DMSO, specific DMSO degradation rates under aerobic and anoxic conditions were both lower than 4 mg DMSO/g VSS-hr. Under anaerobic conditions, the specific DMSO degradation rate was estimated to be 14 mg DMSO/g VSS-hr, which was much higher than those obtained under aerobic and anoxic conditions. The optimum specific MEA and TMAH degradation rates were obtained under aerobic conditions with values of 26.5 mg MEA/g VSS-hr and 17.3 mg TMAH/g VSS-hr, respectively. Compared to aerobic conditions, anaerobic biodegradation of MEA and TMAH was much less significant with values of 5.6 mg MEA/g VSS-hr and 0 mg TMAH/g VSS-hr, respectively. In summary, biological treatment of TFT-LCD wastewater containing DMSO, MEA, and TMAH is feasible, but appropriate conditions for optimum biodegradation of DMSO, MEA, and TMAH are crucial and require carefully operational consideration.
Assuntos
Resíduos Industriais , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da Água/isolamento & purificação , Purificação da Água/métodos , Biodegradação Ambiental , Reatores Biológicos/microbiologia , Dimetil Sulfóxido/isolamento & purificação , Dimetil Sulfóxido/metabolismo , Etanolamina/isolamento & purificação , Etanolamina/metabolismo , Compostos de Amônio Quaternário/isolamento & purificação , Compostos de Amônio Quaternário/metabolismo , Taiwan , Poluentes Químicos da Água/metabolismoRESUMO
An effective method for removing dimethylsulfoxide (DMSO) from wastewater, based on the mild selective catalytic sulfoxidation with H(2)O(2), is proposed in this study. The catalysts are W-containing layered double hydroxides (LDH), and they were obtained by ionic exchange of the nitrate anions from MAl-LDH precursors (M=Mg(2+) or Zn(2+)) with both WO(4)(2-) and W(7)O(24)(6-) species. Results of X-ray diffraction (XRD), N(2) adsorption, thermal gravimetric analysis (TGA), diffuse reflectance ultraviolet spectroscopy (DRUV) and Raman spectroscopy confirmed the formation of the W-oxospecies inside the interlayer space and the modification of the textural properties upon the exchange process. All catalysts showed very good activity and stability in the DMSO conversion into dimethylsulfone with dilute H(2)O(2) aqueous solution, at low temperatures (20-50 degrees C). The efficiency of the H(2)O(2) was higher than 95% and the behaviour of the water as solvent was very close to that of the organic solvents (ethanol, acetonitrile, 1,4-dioxane).
Assuntos
Dimetil Sulfóxido/isolamento & purificação , Recuperação e Remediação Ambiental/métodos , Eliminação de Resíduos Líquidos/métodos , Hidróxido de Alumínio/química , Catálise , Combinação de Medicamentos , Peróxido de Hidrogênio/química , Hidróxido de Magnésio/química , Oxirredução , Compostos de Tungstênio/química , Difração de Raios XRESUMO
BACKGROUND: Umbilical cord blood (UCB) is an important source of hematopoietic stem and progenitor cells (HSC/HPC) for the reconstitution of the hematopoietic system after clinical transplantation. Cryopreservation of these cells is critical for UCB banking and transplantation as well as for research applications by providing readily available specimens. The objective of this study was to optimize cryopreservation conditions for CD34+ HSC/HPC from UCB. METHODS: Cryopreservation of CD34+ HSC/HPC from UCB after mononuclear cell (MNC) preparation was tested in a research-scale setup. Experimental variations were concentration of the cryoprotectant, the protein additive and cell concentration. In addition, protocols involving slow, serial addition and removal of DMSO were compared with standard protocols (fast addition and removal of DMSO) in order to avoid osmotic stress for the cryopreserved cells. Viability and recoveries of MNC, CD34+ cells and total colony-forming units (CFU) were calculated as read-outs. In addition, sterility testing of the collected UCB units before further processing was performed. RESULTS: The optimal conditions for cryopreservation of CD34+ HPC in MNC preparations were 10% DMSO and 2% human albumin at high cell concentrations (5 x 10(7) MNC/mL) with fast addition and removal of DMSO. After cryopreservation using a computer-controlled freezer, high viabilities (89%) and recoveries for CD34+ cells (89%) as well as for CFU (88%) were observed. Microbial contamination of the collected UCB samples was reduced to a rate of 6.4%. DISCUSSION: Optimized cryopreservation conditions were developed for UCB MNC in respect of the composition of the cryosolution. In addition, our results showed that fast addition of DMSO is essential for improved cryopreservation and post-thaw quality assessment results, whereas the speed of DMSO removal after thawing has little influence on the recoveries of CD34+ cells and CFU.
Assuntos
Criopreservação/métodos , Sangue Fetal/citologia , Células-Tronco Hematopoéticas/citologia , Antígenos CD34/análise , Bactérias/isolamento & purificação , Contagem de Células , Separação Celular/métodos , Sobrevivência Celular , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Dimetil Sulfóxido/química , Dimetil Sulfóxido/isolamento & purificação , Células Precursoras Eritroides/citologia , Sangue Fetal/microbiologia , Células-Tronco Hematopoéticas/química , Humanos , Contagem de Leucócitos , Leucócitos Mononucleares/química , Leucócitos Mononucleares/citologia , Albumina Sérica/química , Células-Tronco/citologiaRESUMO
BACKGROUND: The direct transfusion of thawed hematopoietic progenitor cells (HPCs) is associated to transfusion-related side effects that are thought to be dose-dependent on the infused dimethyl sulfoxide (DMSO). Both the effectiveness of a fully automated cell processing device to washing out DMSO and the effects of DMSO elimination over the recovered cells were evaluated. STUDY DESIGN AND METHODS: Twenty cryopre-served peripheral blood HPC bags (HPC apheresis [HPC-A]) were thawed and processed for washing with an automated cell-processing device. Viability, colony-forming units (CFUs), and absolute count of recovered cells were evaluated by flow cytometry immediately after washing as well as at different times after washing and compared with a sample taken just after thawing (control) but maintained at 4 degrees C. DMSO content was measured by high-performance liquid chromatography and the osmolarity with an osmometer. RESULTS: The median recovery of viable total nucleated cells, viable CD34+ cells, and CFU colonies was 89 (range, 74-115), 103 (range, 62-126), and 91 percent (range, 46%-196%), respectively, in the washing group. Recovery of viable CD3+ cells was 97 percent (range, 42%-131%) and CD14+ cells was 82 percent (54%-119%). The percentages of DMSO elimination and osmolarity reduction were 98 (range, 96-99) and 90 percent (range 86%-95%), respectively. Moreover, elimination of the cryoprotectant improved CFU count, viability, and cell recoveries along the time when compared with the control group. CONCLUSION: Washing out DMSO in thawed HPC-A by use of this approach is safe and efficient in terms of recovery and viability of nucleated and progenitor cells. Additionally, the removal degree of DMSO is very high and therefore might ameliorate the transfusion-related side effects.
Assuntos
Crioprotetores/isolamento & purificação , Dimetil Sulfóxido/isolamento & purificação , Transplante de Células-Tronco Hematopoéticas , Antígenos CD34/análise , Remoção de Componentes Sanguíneos , Criopreservação , HumanosRESUMO
It has been hypothesized that, in addition to freezing injury, some damage to platelets may result from the cell packing that occurs during removal of the cryoprotectant. This study examined DMSO removal by fluid exchange across hollow-fiber (HF) filters as an alternative to centrifugation. The DMSO solution with or without cell suspension was passed once through the filter. The optimum exchange during unloading of DMSO was determined by varying the flow rates in the external and internal compartments of the HF filter. Initially, buffered solutions of a 5% DMSO solution in the absence of platelets were pumped into the fibers and exchanged against PBS. The residual DMSO was determined by osmometry. The exchange of DMSO across the membrane was flow dependent and also influenced by the chemical nature of the HF fibers. No protocol using a reasonable rate flow through the fibers removed more than 95% of the DMSO in a single pass. The optimum protocol was achieved with polysynthane fibers with an internal flow rate of approximately 20 mi/min and an external flow rate of 100 ml/min. Subsequently, frozen/thawed platelet concentrates in DMSO were washed using centrifugation and compared to the HF filtration method. Platelet quality was assayed by flow cytometry, cell count, morphology and osmotic stress test. Both filtration and centrifugal washing techniques resulted in comparable morphological scores and numbers of discoid cells. When agents reducing platelet activation were added, platelet quality was improved after washing by either technique. The lower platelet osmotic response with HF filtration than with centrifugation while using activation inhibitors was attributed to the remaining amount of the inhibitors. All other parameters tested were similar. The expression of CD62P was equivalent with both techniques, and centrifugation did not activate platelets more than filtration contrary to what was originally anticipated. In conclusion, platelet quality was comparable after washing by either technique but hollow fiber filtration does remove cryoprotectant more rapidly than does centrifugation.
Assuntos
Plaquetas , Preservação de Sangue , Dimetil Sulfóxido/isolamento & purificação , Filtração/métodos , Sobrevivência Celular , Centrifugação , Criopreservação , Filtração/instrumentação , Filtração/normas , Humanos , Membranas Artificiais , Inibidores da Agregação Plaquetária , Transfusão de PlaquetasRESUMO
Infusion of dimethylsulfoxide (DMSO) contained in cryopreserved and thawed hematopoietic stem cell (HSC) grafts is frequently associated with mild or moderate adverse reactions, and occasionally with more severe events including neurological symptoms. The severity of these complications is related to the amount of residual DMSO. We evaluated a recently available, closed, automated and 'cgmp (current good manufacturing practice) compliant' device (CytoMate) for its ability to wash out DMSO at the expense of a limited loss of viable CD34(+) cells. A total of 16 procedures were carried out with 39 blood HSC bags intended for destruction. Mean amounts of DMSO for each cellular product (one, two or three bags) were between 12.2 and 39.6 g before thawing; after the washing procedure, residual DMSO quantities were between 0.1 and 3.7 g. When set up to reproducibly allow for a more than 96% elimination of DMSO, processing of thawed cells with the CytoMate cell processor resulted in a mean recovery of viable total cells, CD34(+) cells and lymphocyte subsets above 60%. We conclude that this simple and efficient washing technique is suitable for routine processing of HSC grafts. Clinical studies will demonstrate whether a reduction in the incidence of adverse effects associated with DMSO infusion is observed.
Assuntos
Criopreservação/instrumentação , Dimetil Sulfóxido , Transplante de Células-Tronco Hematopoéticas/instrumentação , Preservação de Tecido/instrumentação , Automação , Remoção de Componentes Sanguíneos/instrumentação , Remoção de Componentes Sanguíneos/métodos , Sobrevivência Celular , Criopreservação/métodos , Crioprotetores/análise , Crioprotetores/isolamento & purificação , Dimetil Sulfóxido/análise , Dimetil Sulfóxido/isolamento & purificação , Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas , Humanos , Contagem de Linfócitos , Reprodutibilidade dos Testes , Preservação de Tecido/métodos , Transplante AutólogoRESUMO
With water as the elution solvent, zwitterionic solutes and polyols were retained on HPLC columns, more than was water, by totally hydrophobic packing materials. Relative retentions were systematically affected by oxygen functional groups in the packing material, explicable as specific retention of water. Reproducible elution sequences of 20 solutes at a variety of hydrophobic surfaces (aromatic and both long- and short-alkyl aliphatic surfaces) showed there is a general process, consistent with interactions with hydration water at the surface having solvent properties distinct from bulk water. Early eluting solutes included glycine, sarcosine and taurine. Glycine betaine followed both these and N,N-dimethylglycine. The natural betaines propionobetaine and dimethylsulfoniopropionate also preceded glycine betaine. Dimethylsulfoxide was strongly retained, as (to a lesser extent) was proline betaine. Polyols eluted in the sequence sorbitol, trehalose, glycerol. Changes in the chemical nature of the surface or base material affected relative retentions of water and solutes. The presence of hydrogen-bonding functions increased retention of polyols, as well as water, relative to zwitterionic solutes. Specific effects retention, constraining models based on the formation of low-density water.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Polímeros/isolamento & purificação , Óxido de Alumínio , Betaína/isolamento & purificação , Fenômenos Químicos , Físico-Química , Dimetil Sulfóxido/isolamento & purificação , Estabilidade de Medicamentos , Glicina/isolamento & purificação , Ligação de Hidrogênio , Íons , Modelos Químicos , Concentração Osmolar , Polímeros/química , Pressão , Proteínas/química , Proteínas/isolamento & purificação , Sarcosina/análogos & derivados , Sarcosina/isolamento & purificação , Dióxido de Silício , Soluções , Solventes , Taurina/isolamento & purificação , ÁguaRESUMO
A method for separating low molecular cryoprotectiva from freeze-conserved erythrocyte- and thrombocyte-concentrates by exclusion chromatography has been described. A new vesicular packing material has been used. Only 25 to 30 minutes are necessary in order to separate glycerol respectively dimethylsulphoxide (DMSO) completely from the cells. 86.5% of the erythrocytes and 75.4% of the thrombocytes were recovered after the separation process.
Assuntos
Plaquetas/citologia , Preservação de Sangue/métodos , Crioprotetores/isolamento & purificação , Eritrócitos/citologia , Cromatografia em Gel/métodos , Criopreservação , Dimetil Sulfóxido/isolamento & purificação , Glicerol/isolamento & purificação , HumanosRESUMO
A method for quantitative determination of gentamicin and dimethylsulfoxide (DMSO) in "Gentaplast", a film-forming aerozol preparation was developed. Gentamicin was determined microbiologically after extraction. DMSO was determined by titration with the use of the Redox method. The results of determination of gentamicin and DMSO in "Gentaplast" were statistically treated.