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At least 253 records · Page 14Linked to original sources
PubMed1997

Comparison of the effects of red cell separation and ultrafiltration on heparin concentration during pediatric cardiac surgery.

OBJECTIVE: To determine the effects of red cell separation and ultrafiltration on heparin concentration. DESIGN: Prospective study. SETTING: University-affiliated, pediatric medical center. PARTICIPANTS: Thirty-one children undergoing cardiac surgery. INTERVENTIONS: Blood sampled for heparin concentration and coagulation tests. MEASUREMENTS AND MAIN RESULTS: Thirteen infants under-went modified veno-venous ultrafiltration (UF) after cardiopulmonary bypass (CPB). In addition, residual blood in the CPB circuit was hemoconcentrated by UF and reinfused (UF group). Heparin concentration increased from 2.0 +/- 0.6 to 2.5 +/- 0.8 U/mL, following modified UF; while activated coagulation time (ACT) decreased from 701 +/- 177 to 627 +/- 107 seconds. Heparin concentration of CPB circuit residual increased from 1.9 +/- 0.7 to 3.1 +/- 1.0 U/mL. In 18 children (older than 1 year old), the residual blood in the CPB circuit was hemoconcentrated by cell separation (CS) and reinfused (CS group). Heparin concentration of CPB circuit residual decreased from 2.6 +/- 0.6 to 0.3 +/- 0.2 U/mL. After reinfusion, patient heparin concentration remained unchanged at < 0.05 U/mL. Thrombin time increased from 28 +/- 6 to 48 +/- 29 seconds and did not correlate with H. CONCLUSIONS: The plasma concentration of heparin increased after veno-venous modified UF of the patient. Heparin concentration also increased after UF of residual CPB circuit blood. In contrast, circuit blood hemoconcentrated by CS contained minimal heparin, and, when infused, did not increase patient's heparin concentration. ACT and thrombin time did not correlate with heparin concentration.

Blood Transfusion↗
PubMedDate not supplied

Neutrophil transfusion. In vivo function of neutrophils collected using cell separators.

Effective transfusion therapy depends, in part, upon the ability of the transfused neutrophils to function normally in the recipient. To evaluate the in vivo function of neutrophils obtained with two widely used newer cell separators (Fenwal CS-3000 and IBM 2997), neutrophil concentrates from 12 normal subjects were labeled in vitro with 3H-diisopropylfluorophosphate, reinfused into the donor, and blood kinetics and skin chamber accumulation of the labeled cells were measured. Results were compared with previously obtained data (referred to as "normal") using neutrophils collected by phlebotomy. Blood kinetic measurements of cells obtained with the CS-3000 were normal, whereas blood recovery of cells obtained with the 2997 was decreased moderately. The percent of infused neutrophils localizing to the skin chamber was 0.101 and 0.044 for CS-3000 and 2997 cells, respectively; neither value was significantly different from normal (0.066). Thus, neutrophils collected using either of these cell separators function normally or nearly so in vivo.

Blood Transfusion↗
PubMed1979

In situ lymphoid cells of mouse mammary tumors. IV. Comparison of functional activity of lymphoid cells separated from mammary tumors to that of spleen and lymph node cells of tumor-sensitized mice.

Lymphoid cells isolated form several types of mouse mammary tumors are capable of stimulating tumor cell growth or survival in MCT assays. Lymph node and spleen cells of mice bearing such a tumor are specifically cytotoxic to the tumor cells. Surgical removal of the tumor is followed in 4 to 7 days by the appearance of stimulatory capacity in spleens and lymph nodes. By day 10, cytotoxic cells specific for the sensitizing tumor are again detected. These reach a peak on day 13. By day 17 no reactivity is detectable. The functional distribution of tumor-reactive lymphoid cells is different between tumor masses and peripheral lymphoid organs.

Animals↗
PubMed2006

Autologous bone-marrow mononuclear cell implantation in patients with severe lower limb ischaemia: a comparison of using blood cell separator and Ficoll density gradient centrifugation.

Different studies have provided evidence that implantation of bone-marrow mononuclear cells (BM-MNC) into ischaemic limbs can improve tissue vascularization. Based on these results we performed a pilot study in patients with critical lower limb ischaemia to assess efficacy and safety of implantation of autologous BM-MNC. The amount and efficacy of BM-MNC purified either by an automated method or by a manual procedure were compared. Twelve patients with severe unilateral lower limb ischaemia were entered into this study. They were randomly assigned to be injected with BM-MNC sorted on a blood cell separator or isolated by density gradient on Ficoll-Hypaque. BM-MNC were implanted into the ischaemic legs. Patients were monitored with resting ankle-brachial pressure index (ABI), arterial oxygen saturation (SaO(2)), pain-free walking time and rest pain scale evaluation. The automated and manual methods used for mononuclear cell separation gave results not significantly different. Monitored variables improved in both groups. Improvement of ischaemic condition persisted during 24 weeks follow-up. Limb salvage was achieved in five cases. Our results indicate that BM-MNC implantation into ischaemic limbs is a practical, safe and effective method that may significantly contribute to the management of patients with limb ischaemia. The Ficoll method is a simple and effective procedure for BM-MNC concentration that may be useful, mainly in hospitals without sophisticated facilities.

Aged↗
PubMed1986

Enrichment of human bone marrow aspirates for low-density mononuclear cells using a haemonetics discontinuous blood cell separator.

Isopycnic density floatation centrifugation has been proven to be a suitable technique to enrich bone marrow aspirates for clonogenic cells on a small scale. We have tested a Haemonetics semicontinuous blood cell separator in order to process large volumes of bone marrow with minimal bone marrow manipulation. The efficacy of isopycnic density floatation was tested in a one and a two-step procedure. Both procedures showed a recovery of about 20% of the nucleated cells and 1-2% of the erythrocytes. The enrichment of clonogenic cells in the one-step procedure appeared superior to the two-step enrichment, first separating buffy coat cells. The recovery of clonogenic cells was 70 and 50%, respectively. Repopulation capacity of the low-density cell fraction containing the clonogenic cells was excellent after autologous reinfusion (6 cases) and allogeneic bone marrow transplantation (3 cases). Fast enrichment of large volumes of bone marrow aspirates with low-density cells containing the clonogenic cells by isopycnic density floatation centrifugation can be done safely using a Haemonetics blood cell separator.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2001

Collection of MNCs with two cell separators for adoptive immunotherapy in patients with stage IV melanoma.

BACKGROUND: MNCs for adoptive immunotherapy may be collected by leukocytapheresis with a cell separator. STUDY DESIGN AND METHODS: Six healthy cytapheresis donors donated two MNC concentrates on a cell separator (AS.TEC 204, Fresenius): one on the standard MNC program and one on a modified MNC program with reduced centrifuge velocity that leads to a lower platelet contamination. Seventeen patients with malignant melanoma donated 26 MNC concentrates: 5 on the AS.TEC 204 MNC program, 9 on the modified AS.TEC 204 MNC program, and 12 on another modified MNC program (Spectra, COBE). RESULTS: In the course of cultivation of MNCs to dendritic cells (DCs), the donor MNC concentrates with the lower platelet contamination (475 +/- 85 x 10(9)/L) had a significantly higher relative DC yield (low platelet contamination: 3.9 +/- 1.6% of the plated cells; high platelet contamination: 2.5 +/- 1.8% of the plated cells; p = 0.019) than the concentrates with the higher platelet contamination (2364 +/- 448 x 10(9)/L). No significant difference was found in the yields of MNCs and CD14+ cells in the three protocols used for the collection of MNCs from patients with melanoma. The components obtained by the standard AS.TEC 204 MNC program had a significantly higher platelet contamination (1768 +/- 994 x 10(9)/L) than the components obtained by the modified AS.TEC MNC program (360 +/- 98 x 10(9)/L; p<0.05) and the modified Spectra MNC program (636 +/- 266 x 10(9)/L); p<0.05). Because of the low number of investigated components, no significant difference in the DC yield of the three protocols could be detected (mean DC yield after cultivation: 746 +/- 429 x 10(6)). CONCLUSION: A high platelet contamination of MNC concentrates intended for adoptive immunotherapy can lead to a significant impairment of the DC yield after cultivation. Both the modified AS.TEC 204 and the modified Spectra MNC programs are well suited for collecting MNC concentrates with high MNC yields and low platelet contamination from patients with malignant melanoma.

Adult↗
PubMed1974

Studies on dispersed pancreatic exocrine cells. I. Dissociation technique and morphologic characteristics of separated cells.

A procedure for dissociation of the guinea pig pancreas into individual cells is described which employs enzymatic digestion with pure collagenase, chymotrypsin, and hyaluronidase, utilizes an interposed chelation of divalent cations by EDTA, and is terminated by gentle shearing. Yields of cells are 50-60%, based on DNA recovered. The population comprises approximately 95% exocrine cells, the remainder consisting of endocrine, duct, and vascular endothelial cells. The exocrine cells, though spherical, retain the structural attributes of their in situ counterparts, including differentiation of the plasmalemma into zones corresponding to the former apical and basal plasmalemma, polarized distribution of organelles indicated by fields of zymogen granules in the cytoplasm underlying the former apex, central location of the Golgi complex, and placement of the rough endoplasmic reticulum and nucleus in the former basal pole of the cell. Electron microscope study of the effects of individual treatments used during dissociation indicates that digestion of basement membrane and collagen is solely due to collagenase activity and that separation of desmosomes (and possibly of zonulae adherentes) results only from exposure to low [Ca(++)] and EDTA and is not effected by the enzymes used. Gap junctions are resistant to enzymes and EDTA; tight junctions resist enzyme treatment but undergo rearrangement upon exposure to EDTA. Both junctions require mechanical shear for complete cell separation. Neither chymotrypsin nor hyaluronidase produces visible alterations in stromal or junctional elements. Dissociation requires the concerted action of enzymes, chelation of divalent cations, and mechanical shear, since the individual treatments are alone ineffective.

Animals↗
PubMed1994

[Second multicenter study of thrombocytapheresis using the Fresenius AS104 blood cell separator and ring study for quality control of cell counts. Members of the Preparative and Therapeutic Hemapheresis Section of DGTI].

The aim of the 2nd Multicenter study was to evaluate the separation protocol software version V 4.61 of the Fresenius AS 104 cell separator, for separation efficiency, WBC contamination and yield's deviation from prediction. Plateletpheresis data from 12 hemapheresis centers, using identical apheresis protocols and cell counting methods, were registered and statistically analyzed. Additionally, the counting methods of the centers were controlled by a ring study with biweekly cell counts. To get a comparison the apheresis data, which were dependent of the center effects, were corrected by the systematical deviation found in the quality control from the ring study. The results of 935 runs are 47.3 +/- 8.1% for the separation effectivity. 7.2% median deviation from predicted yield, whereby 90% of all runs deviated less than +/- 22% from predicted yields. 50% of products had a WBC contamination below 6 x 10(6), 99% below 5 x 10(7).

Blood Component Removal↗
PubMed2003

Novel method for continuous cell separation by density gradient centrifugation: evaluation of a miniature separation column.

A compact bench-top model of the centrifuge enables continuous cell separation based on density differences. The apparatus holds a small separation disk equipped with a circular channel (8 mL capacity) separated by a septum. A set of isotonic Percoll media with different densities is continuously introduced at one terminal and collected from the other. Under a centrifugal force field, cell suspension introduced into the proximal portion of the channel results in continuous separation of cells according to their densities. The performance of the apparatus was demonstrated with the separation of human buffy coat containing nucleated cells (>10(8)) among a large population (10(10)) of RBC. The results indicated that the method is capable of separating a large number of nucleated cells, with minimum damage, for a few hours of operation wherein neutrophils are well resolved from lymphocytes. The method may be applied to other types of samples including cord blood, blood from small animals, cultured cells, pancreatic beta cell islets, malaria parasites, sperm cells, etc.

Blood Cell Count↗
PubMed1994

Correlation of Pectin Methylesterase Activity in Root Caps of Pea with Root Border Cell Separation.

We tested predictions of the hypothesis that pectin methylesterase in the root cap plays a role in cell wall solubilization leading to separation of root border cells from the root tip. Root cap pectin methylesterase activity was detected only in species that release large numbers of border cells daily. In pea (Pisum sativum) root caps, enzyme activity is correlated with border cell separation during development: 6-fold more activity occurs during border cell separation than after cell separation is complete. Higher levels of enzyme activity are restored by experimental induction of border cell separation. A corresponding increase in transcription of a gene encoding root cap pectin methylesterase precedes the increase in enzyme activity. A dramatic increase in the level of soluble, de-esterified pectin in the root tip also is correlated with pectin methylesterase activity during border cell development. This increase in acidic, de-esterified pectin during development occurs in parallel with a decrease in cell wall/apoplastic pH of cells in the periphery of the root cap.

Journal Article↗
PubMed1978

Magnetic microspheres prepared by redox polymerization used in a cell separation based on gangliosides.

A facile method is described for making magnetic microspheres that bind specifically to cell surfaces, in order to separate cells magnetophoretically. Control over the sizes of the spheres is effected by using their magnetic cores as part of a redox polymerization system. The use of the microspheres is demonstrated with a separation involving C-1300 neuroblastoma cells, 10% of which express the ganglioside GM1 in their membranes. The GM1-containing cells were separated with better than 99% purity, while the deficient cells were obtained at least 98% pure. The separation, which was carried out under sterile conditions, required only 6 minutes.

Cell Line↗
PubMed2004

Collection of peripheral progenitor cells: a comparison between Amicus and Cobe-Spectra blood cell separators.

The authors compared the efficiency of two different blood cell separators (Amicus and Cobe-Spectra) in collecting peripheral blood progenitor cells for autologous or homologous transplantation. A total number of 129 procedures were performed, 36 with Spectra, 93 with Amicus. There was no difference between Spectra and Amicus efficiencies for CD34+ cell collection (46.685% vs 46.235%; p=n.s) but the platelet efficiencies were 17.31% and 12.54% respectively (p=0.04) and, if autologous and allogeneic collections were considered separately, a marked difference resulted in allogeneic platelet efficiency between 6 Spectra and 23 Amicus procedures (26.83% vs 8.68%, p=0.0004). The authors were able to demonstrate that in 70 Amicus autologous collections there was a different platelet efficiency, if peripheral count was considered: 12 procedures performed with a platelet count > 100 x 10(9)/l had a very low efficiency (6.86%), but this value increased if platelet count lowered (13.02% if between 100 and 50 x 10(9)/l, 22.63% if between 50 and 0 x 10(9)/l, 23 and 35 procedures respectively). The study is preliminary and the number of collections is little, but the overall data suggest that Spectra (AutoPBSC, V 6.0) and Amicus separators have the same efficiency for collecting CD34+ cells while Amicus procedures have a very low platelet contamination, especially with donors.

Antigens, CD34↗
PubMed2004

Peripheral blood cell separation through surface-modified polyurethane membranes.

Cell separation from peripheral blood was investigated using surface-modified polyurethane (PU) membranes with different functional groups. Both red blood cells and platelets could pass through unmodified PU and PU-SO(3)H membranes, whereas the red blood cells preferentially passed through PU-N(C(2)H(5))(2) and PU-NHC(2)H(4)OH membranes. The permeation ratio of T and B cells was <25% for the surface-modified and unmodified PU membranes. CD34(+) cells have been recognized as various kinds of stem cells including hematopoietic and mesenchymal stem cells. The adhesiveness of CD34(+) cells on the PU membranes was found to be higher than that of red blood cells, platelets, T cells, or B cells. Overall, the adhesiveness of blood cells on the PU membranes increased in the following order: red blood cells </= platelets < T cells </= B cells < CD34(+) cells. Treatment of PU-COOH membranes with a human albumin solution to detach adhered blood cells, allowed recovery of mainly CD34(+) cells in the permeate, whereas both red blood cells and platelets could be isolated in the permeate using unmodified PU membranes. The PU membranes showed different permeation and recovery ratios of specific cells depending on the functional groups attached to the membranes.

Antigens, CD↗
PubMed1977

Plateletpheresis using the haemonetics model 30 cell separator.

Plateletpheresis using the Haemonetics Model 30 cell separator results in a mean collection of 5.5 X 10(11) platelets. This product contains substantial numbers of erythrocytes and mononuclear cells which can be effectively removed by centrifugation but with a 23% loss of platelets as well. The mean decrement in donor's platelet count during the procedure, which is 78,000/mm3, is not commensurate with the number of platelets collected suggesting donor platelet mobilization. Platelets collected by this technic, when transfused into unselected recipients, give platelet increments comparable to platelets produced in the standard batch manner.

ABO Blood-Group System↗
PubMed1988

Molecular design of materials for cell separation.

There has been a strong demand in biomedical sciences to isolate viable cell populations with high yield and purity. An important facet of this work was to develop new polymeric adsorbent for the separation of lymphocyte subpopulations. Based on our strategy of separating cells through their differential ionic affinity toward multiphase-structured adsorbent with ionically derivatized microdomains, a series of poly(2-hydroxyethyl methacrylate)/polyamine graft copolymers (HA copolymers) was prepared. HA copolymer columns were found to show specific adsorption affinity toward B lymphocytes, and allows for separation of B and T lymphocytes in high yield and purity with a short operating time. Separation mechanism involved in the resolution of B and T lymphocytes by HA copolymer column is discussed in this paper. Further, photo-induced desorption of cells from the adsorbent derivatized with photo-responsive functional group (azobenzene group) was demonstrated to emphasize the feasibility of photo-regulated chromatography as a novel tool in cell separation technology.

B-Lymphocytes↗
PubMed1987

Side-effects of cytapheresis with cell separators.

The rate of procedure-related side-effects was evaluated in 11,895 cytaphereses performed with a Haemonetics 30 (H-30) cell processor (2,486 procedures) or with a Haemonetics V-50 (H-50) cell processor (6,813 procedures) or with the Fenwal CS-3000 cell processor (2,596 procedures). The overall rate of side-effects was 7.41% (absolute number 881). There was a significant difference between female and male donors concerning the frequency of hypotension episodes and citrate toxicity related reactions. There were also distinct differences between the single cell separator types concerning the frequency and type of side effects. Hypotension episodes and citrate-related reactions were observed most frequently during cytapheresis with the H-30 (7.65 and 2.96% respectively). Venous access and technical problems appeared more often when the H-50 or the CS-3000 were used. Thus both the H-50 as well as the CS-3000 cell separators allow safe cytapheresis with a low number of side-effects.

Austria↗
PubMed1999

Controlled shear filtration: A novel technique for animal cell separation.

A novel rotary microfiltration technique specifically suited for the separation of animal cells has been developed. The concept allows the independent adjustment of wall shear stress, transmembrane pressure, and residence time, allowing straightforward optimization of the microfiltration process. By using a smooth, conically shaped rotor, it is possible to establish a controlled shear field in which animal cells experience a significant hydrodynamic lift away from the membrane surface. It is shown in preliminary experiments that shear-induced cell-rupture speeds up membrane clogging and that cell debris poses the most significant problem in harvesting of BHK cell cultures by dynamic microfiltration. However, a threshold value of shear stability exists which depends on the frequency of passing the shear field, the residence time in the shear field, as well as on cell status. By operating close to this threshold value, cell viability can be maintained while concentration polarization is efficiently minimized. By applying this concept, it is possible to attain flux rates several times higher compared to conventional crossflow filtration. Controlled shear filtration (CSF) can be used for batch harvesting as well as for cell retention in high cell density systems. In batch harvesting of hIL-2 from rBHK cell culture, a constant flux rate of 290 L h-1 m-2 has been adjusted without indication of membrane clogging or fouling.

Animals↗