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At least 19 recordsLinked to original sources
PubMed1978

Cell separation, cell differential and granulocyte colony frequency in polycythemia vera.

In seven patients with polycythemia vera, the agar colony growth of bone marrow total nucleated cell suspensions and of the cell fractions obtained with an albumin discontinuous density gradient were studied. In one patient, the density distribution of colony-forming units in culture (CFUc) before and after alkylating treatment was evaluated and cell differentials on smears obtained from each density fraction were determined. A high percentage of low density CFUc compared with the total CFUc population and with that in the normal control subjects in the same density fractions was observed.

Bone Marrow↗
PubMed2005

Role of septins and the exocyst complex in the function of hydrolytic enzymes responsible for fission yeast cell separation.

Cell separation in Schizosaccharomyces pombe is achieved by the concerted action of the Eng1 endo-beta-1,3-glucanase and the Agn1 endo-alpha-1,3-glucanase, which are transported to the septum and localize to a ringlike structure that surrounds the septum. The requirements for the correct localization of both hydrolases as a ring were analyzed using green fluorescent protein fusion proteins. Targeting to the septum required a functional exocyst, because both proteins failed to localize correctly in sec8-1 or exo70delta mutants, suggesting that Agn1 and Eng1 might be two of the cargo proteins present in the vesicles that accumulate in exocyst mutants. Septins and Mid2 were also required for correct formation of a ring. In their absence, Eng1 and Agn1 were found in a disk-like structure that spanned the septum, rather than in a ring. Even though septin and mid2delta mutants have a cell separation defect, the septum and the distribution of linear beta-1,3-glucans were normal in these cells, suggesting that mislocalization of Eng1 and Agn1 might be the reason underlying the failure to separate efficiently. Thus, one of the functions of the septin ring would be to act as a positional marker for the localization of hydrolytic proteins to the medial region.

Cytokinesis↗
PubMed1994

An experimental model of affinity cell separation.

Cell affinity separations are based on the selective attachment of cell phenotype using antibody or lectins specific for cell surface markers. The major physicochemical factors which influence ligand-mediated cell adhesion dynamics and the efficiency of cell affinity separation have been examined. Uniform cell detachment forces were generated with a parallel-plate flow cell (plate separation 100 microns, surface area 3 cm2). Hydrodynamic shear stress was used to measure cell adhesion strength and to separate cells on the basis of surface affinity. Human cell lines grown in tissue culture were separated on a flat derivatised glass immunoadsorbent which formed the floor of the flow chamber. Flow-cell residence time, detachment shear stress, temperature, and ligand density were shown to influence cell attachment probability. An understanding of the physical basis of ligand-mediated cell adhesion provided a rationale for optimisation of affinity cell separation. At room temperature attachment of positive cells was rapid (< 2 min) and adhesion strength was directly related to immunoadsorbent ligand density. Purity and recovery of enriched fractions were dependent on the separation shear stress and could be optimised using this parameter. Enrichment factors were greater than 100-fold, with at least 90% of positive cells recovered in enriched fractions. Enrichment purity and yields did not decline at higher loading densities (10(5) cells/cm2). Selective immunoadsorbent surface chemistry is a prerequisite for efficient affinity cell separation. Purity and recovery may be optimised by fractionating enriched and depleted cell populations with uniform fluid shear stress.

Antibody Affinity↗
PubMed2004

Role of the alpha-glucanase Agn1p in fission-yeast cell separation.

Cell division in the fission yeast Schizosaccharomyces pombe yields two equal-sized daughter cells. Medial fission is achieved by deposition of a primary septum flanked by two secondary septa within the dividing cell. During the final step of cell division, cell separation, the primary septum is hydrolyzed by an endo-(1,3)-beta-glucanase, Eng1p. We reasoned that the cell wall material surrounding the septum, referred to here as the septum edging, also must be hydrolyzed before full separation of the daughter cells can occur. Because the septum edging contains (1,3)-alpha-glucan, we investigated the cellular functions of the putative (1,3)-alpha-glucanases Agn1p and Agn2p. Whereas agn2 deletion results in a defect in endolysis of the ascus wall, deletion of agn1 leads to clumped cells that remained attached to each other by septum-edging material. Purified Agn1p hydrolyzes (1,3)-alpha-glucan predominantly into pentasaccharides, indicating an endo-catalytic mode of hydrolysis. Furthermore, we show that the transcription factors Sep1p and Ace2p regulate both eng1 and agn1 expression in a cell cycle-dependent manner. We propose that Agn1p acts in concert with Eng1p to achieve efficient cell separation, thereby exposing the secondary septa as the new ends of the daughter cells.

Amino Acid Sequence↗
PubMed1991

[Stem cell separation with different cell separators].

For some years, there has been an increasing success in transplanting peripheral blood stem cells (PBSC) instead of autologous bone marrow in patients suffering from different malignancies. While collecting PBSC for autologous transplantation, we compared four different separation techniques and three different cell separators (COBE Spectra, Fresenius AS 104, Haemonetics V50) routinely used for platelet production. Our results suggest that continuous flow separators seem to have some advantage over discontinuous flow machines in harvesting PBSC.

Blood Cell Count↗
PubMed1991

[Optimizing cell separation with the Cell Separator AS-106].

Seventeen different separation protocols based on more than 1100 thrombocytaphereses were tested. Twelve of these are reported on in this study. It became apparent that the most important variable is the centrifugation speed (1900 rpm at a blood flow of 50 ml/min). This made it possible to collect 3.5 to 3.8 x 10(11) thrombocytes (approx. 60% extraction efficiency) from 3.15 l blood. Further optimization of this procedure is desirable, since 10% of the thrombocytes still remain in the separation chamber. However, our optimization work was successful as far as the reduction of leukocyte and erythrocyte contamination is concerned, which ultimately resulted in leukocyte and erythrocyte contamination of about 1 x 10(7).

Blood Component Transfusion↗
PubMed1999

Advances in cell separation: recent developments in counterflow centrifugal elutriation and continuous flow cell separation.

Cell separation by counterflow centrifugal elutriation (CCE) or free flow electrophoresis (FFE) is performed at lower frequency than cell cloning and antibody-dependent, magnetic or fluorescence-activated cell sorting. Nevertheless, numerous recent publications confirmed that these physical cell separation methods that do not include cell labeling or cell transformation steps, may be most useful for some applications. CCE and FFE have proved to be valuable tools, if homogeneous populations of normal healthy untransformed cells are required for answering scientific questions or for clinical transplantation and cells cannot be labeled by antibodies, because suitable antibodies are not available or because antibody binding to a cell surface would induce the cell reaction which should be investigated on purified cells or because antibodies bound to the surface hamper the use of the isolated cells. In addition, the methods are helpful for studying the biological reasons for, or effects of, changes in cell size and cellular negative surface charge density. Although the value of the methods was confirmed in recent years by a considerable number of important scientific results, activities to further develop and improve the instruments have, unfortunately, declined.

Animals↗
PubMed1991

[Tumor cell separation by cell saver and membrane filter passage].

Definite suspensions of malignant cells from three human tumor cells lines (bladder, prostate and renal cell carcinom) were passed through a cell saver (Althin Mediplast) and a leucozyte removal filter (PALL RC 100) under standard conditions. The examination of the solutions did not detect any malignant cells at all. If investigations with malignant cells in the blood will confirm these results, the use of intraoperative autotransfusion in urological tumor surgery would be possible.

Blood Transfusion, Autologous↗
PubMed1984

Responsiveness of human T-lymphocyte subpopulations in autologous mixed-lymphocyte reaction using xenoprotein-free separated cells: autologous reactivity lies chiefly in a low density T-cell fraction.

The responsiveness in the autologous mixed-lymphocyte reaction (AMLR) by human T cells separated using two different methods not involving xenoantigen contract was examined. Although T cells from nylon-wool columns were active in AMLR, T cells separated by a Percoll gradient method responded poorly. Further separation of T cells from nylon-wool columns into low density (TL) and high density (TH) fractions by Percoll revealed that TL was enriched, while TH was depleted, in AMLR responsiveness. This difference could not be accounted for by differences in the helper or suppressor cells in the fractions. Moreover, TH responded well in secondary AMLR. Therefore the T cells reactive in AMLR reside chiefly, although not exclusively, in the low density fraction.

Cell Separation↗
PubMed1990

[Comparison of thrombocyte concentrates of the cell separator AS 104 with preparations of the cell separator CS 3000 (Fenwal)].

Platelet concentrates of the cell separator AS 104 (Fresenius) are compared with those of the cell separator CS 3000 (Fenwal). At each cell separator two platelet preparation protocols and at CS 3000 additionally a WBC preparation protocol was analysed. Platelet counts before and after cellapheresis as well as separation efficiencies show no significant differences between the separation protocols (A, B) and the two cell separators. Platelets of varying volumes show different separation efficiencies. While in WBC concentrates the separation efficiencies of platelets continuously increase from small (2fl) to large (20 fl) platelets, the efficiencies in platelet concentrates decrease in volume range 2 fl-8 fl, and increase in range 8 fl-20 fl. There are no differences between the cell separators. The efficiencies of platelets with 2 fl and 4 fl volumes differ significantly (p less than 0.001) between the protocols A and B of AS 104. After WBC-donation, platelet loss of donors corresponds with platelet yields of the concentrates for the platelet volume range 2 fl-20 fl. Compared with the calculated platelet loss of donors after platelet donation more small platelets with 2 fl-8 fl volumes are found in platelet concentrates. The yields of platelets with 10 fl-20 fl volumes are in accordance with donor's platelet loss.

Blood Transfusion↗
PubMed2006

Oxidative stress by biphenyl metabolites induces inhibition of bacterial cell separation.

Cell-cell separation of a polychlorinated biphenyl (PCB)-degrading bacterium Comamonas testosteroni TK102 was monitored by flow cytometry. When monohydroxy metabolites of biphenyl (BP) (2-hydroxybiphenyl and 3-hydroxybiphenyl) were added to the culture, cell-cell separation of strain TK102 was inhibited at stationary phase. This inhibition was reproduced on non-PCB degrading bacteria such as Pseudomonas putida PpY101 and Escherichia coli MV1184, but was not observed on Pseudomonas aeruginosa PAO1. An opportunistic pathogen, P. aeruginosa PAO1, produces exopolysaccharide, which is known to scavenge damaging chemicals such as reactive oxygen species (ROS). The higher level of ROS and lipid peroxidants were detected in the cells treated by monohydroxybiphenyls. Fat-soluble vitamin E, which is a lipid radical scavenger, maintained bacterial cell separation during monohydroxybiphenyls treatment. Our results demonstrated that intracellular oxidative stress played an important role in the inhibition of bacterial cell separation during BP metabolism. This study shows that metabolites of environmental pollutants, such as monohydroxylated BP, inhibit bacterial cell separation by oxidative stress.

Antioxidants↗
PubMed1994

[Effective erythrocyte exchange by means of a cell separator in sickle cell anemia].

Sickle-cell anemia is one of the most common hemoglobinopathies. The therapy consists of symptomatical measures like giving analgetics, decreasing blood viscosity, and red cell transfusions. We present a cascuistic of a 23-year-old Turkish female patient with a homozygous form of sickle-cell disease, who was treated with analgetics, multiple red blood cell transfusions and Desferal because of severe pain crises. From November 1992 until June 1993 we performed automated red cell exchange transfusions with six fresh, washed and leukocyte-depleted red blood cell units with a continuous flow cell separator (Cobe Spectra). We propose to prefer the exchange with the cell separator to the conventional transfusion therapy by manual exchange or a chronic transfusion program in severe sickle-cell crises.

Adult↗
PubMed1997

The COBE Spectra cell separator is more effective than the Haemonetics MCS-3P cell separator for peripheral blood progenitor cell harvest after mobilization with cyclophosphamide and filgrastim.

BACKGROUND: Peripheral blood is rapidly replacing bone marrow as a source of hematopoietic progenitor cells for autologous transplantation. The advantages of peripheral blood progenitor cell transplantation are enhanced by the ability to collect sufficient progenitor cells to ensure rapid neutrophil and platelet recovery in a single procedure on some cell separators. STUDY DESIGN AND METHODS: A prospective randomized study was undertaken to compare peripheral blood progenitor cell yields from two cell separators (MCS-3P, Haemonetics and Spectra, COBE). Fifteen consecutive patients were mobilized with cyclophosphamide 2 g per m2 (Day 0) and filgrastim 10 micrograms per kg (Days 1-11). Consecutive collections (Day 10, Day 11) were performed with each machine once: patients were randomly assigned to either machine for the initial collection. RESULTS: Collection time was longer on the MCS-3P (p = 0.001), and the volume processed was greater with the Spectra (p < 0.0001). Despite similar nucleated cell yield (p = 0.62), the yield of CD34+ cells (p = 0.001) and colony-forming units-granulocytic-monocytic (p = 0.0001) was significantly higher with the Spectra. The yield of nucleated cells per unit of blood volume processed was higher for the MCS-3P (p = 0.0007), while the CD34+ cell yield (p = 1) and colony-forming units-granulocytic-monocytic yield (p = 1) per unit of blood volume processed were similar for the two machines. The collection of CD34+ cells at levels > 2 x 10(6) per kg (p = 0.063), 5 x 10(6) per kg (p = 0.031), and colony-forming units-granulocytic-monocytic > 1 x 10(5) per kg (p = 0.25) after a single collection was superior for the Spectra. CONCLUSION: The yield of progenitor cells after collection on the Spectra was superior to that achieved with the MCS-3P, because of the larger volume of blood processed per procedure. This would permit more patients to undergo only one collection.

Blood Volume↗
PubMed1976

Membrane surface properties other than charge involved in cell separation by partition in polymer, aqueous two-phase systems.

When aqueous solutions of dextran and of poly-(ethylene glycol) are mixed above certain concentrations, immiscible, liquid two-phase systems are obtained which are useful for separating cells by partition. Some salts partition unequally between the phases, giving rise to an electrostatic potential difference between them. Partition of cells has therefore been thought to depend predominantly on membrane charge. We now report two instances in which membrane charge either does not determine or is not the main determinant of cell partition. (A) Cell partition coefficients in phase systems approaching the critical point (the component concentrations below which a homogeneous solution occurs) increase, even in phase systems in which the phase potential difference is practically zero. Furthermore, in such systems, the partition coefficient of (human) erythrocytes is not reduced by complete removal of sialic acid. (B) Rat and mouse erythrocytes have sizable partition coefficients in a phase system away from the critical point with no potential difference between the phases. Cell surface interaction with the polymers is probably responsible for cell partition in these cases. Partition studies on erythrocytes from nine mammalian species in phases near the critical point with and without electrostatic potential differences reveal major species-specific differences in the membrane charge/noncharge components. A correlation has been found, in phases near the critical point that have essentially no electrostatic potential difference, between partition coefficient and the ratio of poly/monounsaturated fatty acids in the membranes of red cells from different species. Our present results thus provide parameters for the separation of cells by partition in addition to or instead of membrane charge depending on the polymer and salt composition and concentration selected.

Animals↗
PubMed1990

A large-scale magnetic separator for selective cell separations with paramagnetic microbeads.

An improved magnetic separator has been developed for use in large-scale cell separations. This separation method uses paramagnetic microbeads coated with antibodies that selectively bind to target cells. The magnetic separator attracts the microbead-target cell aggregates and holds these aggregates at its surface while the suspending fluid and nontarget cells flow past. The optimum separator design was determined to be two magnetic assemblies in series along with a peristaltic pump. The assemblies consist of neodymium-iron-boron magnet bars sandwiched between steel bars (magnetic pole pieces). The size and pole spacing of the two magnetic assemblies are designed to be different, so that the first assembly, which captures greater than 99.99% of the microbeads, has good magnetic reach-out and a high magnetic holding force at its surface, while the second assembly has an even higher magnetic holding force at its surface. Studies show that the separator can remove 1 x 10(10) microbeads from a suspension of red blood cells processed at a flow rate of 9 ml/min, so that no microbeads are detected in the effluent.

Animals↗
PubMed1986

Synthesis and application of a poly(ethylene glycol)-antibody affinity ligand for cell separations in aqueous polymer two-phase systems.

The possibility of producing biospecific affinity ligands for separating cells in two polymer aqueous phase systems on the basis of cell surface antigens was investigated. Rabbit anti-human erythrocyte IgG was reacted with cyanuric chloride-activated monomethyl poly(ethylene glycol) (PEG) fractions (molecular weights approximately 200, 1900, and 5000) at various molar ratios of PEG to protein lysine groups. The partition coefficient of the protein in a Dextran/PEG two-phase system increased with increasing degree of modification and increasing PEG molecular weight. There was a concomitant loss in ability to agglutinate human erythrocytes. The ability of the modified IgG to bind to a DEAE-cellulose column was almost eliminated by reaction with the PEG 5000, and was decreased to a lesser extent by PEG 1900. This PEG 1900-modified IgG substantially increased the partition of fresh or fixed human erythrocytes into the PEG-rich phase of a suitable phase system, while having no effect on rabbit cell partition. The partition increase could be inhibited by unmodified anti-human red cell IgG but not by nonspecific unmodified human IgG, demonstrating that the ligand effects were specific for the cell type against which the antibody was raised. A mixture of rabbit and human erythrocytes, which ordinarily have very similar partitions in the phase systems used, could be separated on a countercurrent distribution apparatus using the modified IgG. These results demonstrate the feasibility of producing immunologically specific affinity partition ligands for cell separation.

Affinity Labels↗