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At least 217 records · Page 12Linked to original sources

CFD-aided design of a dynamic filter for mammalian cell separation.

In the present work, a rotating disk filter was designed for mammalian cell separation with the aim of avoiding both cell damage and membrane fouling. Different geometric and operational variables of the rotating disk filter were studied using computational fluid dynamics (CFD) by varying rotor radius, rotor angle, membrane-rotor distance, and angular velocity. The combinations of these variables followed a statistical design, so that an analysis of the CFD results provided correlations describing the average shear stress on the membrane surface and the maximum shear stress in the whole module as a function of the variables studied. Based on these correlations, and on the shear resistance levels of Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cell lines, which were investigated using a cone-and-plate viscosimeter, it was possible to determine the geometry and angular velocity that would minimize both cell damage and membrane fouling. After construction, the filter was tested in filtration experiments at increasing permeate fluxes. Cell viability remained >90% for the duration of the experiments (2.5 h), and no indication of fouling was observed. It was shown that the designed dynamic filter is able to effectively avoid both cell damage and membrane fouling, and thus can be used for mammalian cell harvesting and perfusion.

Animals↗

Cell separation: a review.

This review summarizes currently available techniques for cell separation. Techniques that exploit differences in physical properties of cells are widely used but have a number of limitations. Those that are based on differences in surface properties may more readily permit reproducible separation of a functionally homogeneous population of cells. Unfortunately very few techniques achieve separation of cells on the basis of differences in their functional characteristics.

Cell Adhesion↗

Cell separation using fluorescence emission anisotropy.

The emission anisotropy of selected fluorescent probes which interact with cells and their membranes is a sensitive parameter for studying the structural changes associated with different functional states. Such measurements can now be made on individual living cells at rates of up to 103 per second and the cells separated on the basis of the anisotropy function alone or combined with other physical signals using a multiparameter automated computer-controlled cell separator (MACCS). Thus, selection can be on the basis of simple or complex algorithms reflecting the size, macromolecular content, and rotational mobility of cellular components or liganded reporter molecules. Cells isolated in this manner are sterile, viable, and can be used for outgrowth or biochemical studies related to dynamic changes occurring during differentiation or malignant transformation.

Animals↗

Leukocytapheresis: evaluation of a new procedure for the blood cell separator Fresenius AS 104.

For granulocyte separation from peripheral blood with the Fresenius blood cell separator AS 104 a new procedure and a new tubing set were developed and evaluated. Separation is done in cycles of 400 ml of whole blood processed, with continuous blood flow of 50-55 ml/min, speed 750 rpm. HES-450 is added for sedimentation acceleration together with citrate, ratio 1:10. At the end of each cycle granulocytes were pumped out of the separation chamber into the collection container. After processing 13 cycles of 400 ml whole blood/HES-citrate-volume a total yield of 9.20 +/- 2.80 x 10(9) white cells were collected in volumes of 125 ml. 47 +/- 9% of the leukocytes collected were granulocytes, this corresponds to collection of 1.05 +/- 0.09 x 10(9) granulocytes per liter of processed blood. The efficacy of granulocyte collection is 22.5 +/- 5.6%, viability is more than 95%. With this procedure it is possible to collect granulocytes with the blood cell separator Fresenius AS 104 in therapeutical doses.

Blood Component Transfusion↗

[Use of bacterial toximeters with separate cell cultures for continuous water monitoring].

Two commercially available bacterial toxicity monitors are compared. As test organisms pure cultures of Pseudomonas putida are used in both systems. The bacteria are grown continuously in turbidostatic ("Toxalarm", LAR, Berlin, Germany) or chemostatic ("Stiptox-norm", Siepmann und Teutscher, Gross-Umstadt, Germany) regulated cultures in a selective culture medium in nonsterile devices. Toxic substances can be detected by continuously working bacterial respiration tests. Oxygen consumption is the measuring parameter. The bacterial test suspension is mixed continuously in a fixed proportion with air-saturated test water in a measuring cell. The separate culturing of the bacteria and the carrying out of the tests ensures that neither the bacterial culture is endangered by toxic substances nor the sensitivity can be changed by poison adaptation. Results of "Toxalarm"-tests with several chemicals (e.g. Atrazine, Lindane, 2-Nitro-phenol, Sodiumpentachlorophenolate) are presented. The registration of an alarm event (River Rhine) by "Stiptox-norm" is shown.

Animals↗

[Study of changes in the surface of irradiated thymocytes using a method of separating cells in a 2-phase water-polymer system].

The coefficient of cell distribution in a charged biphase water-polymeric system "polyethyleneglycol-dextran" decreased 60 min after irradiation of rat thymus lymphocytes with doses of 1 to 8 Gy. This dose-dependent effect was mainly related to radiosensitive thymocytes of a low fraction of density gradient; it disappeared with the uncharged system and was probably connected with the reduction in the negative electric charge of the irradiated cell surface.

Animals↗

Effect of released lymphocyte proteins on human lymphocytes in vitro. II. Protein synthesis of nylon-wool separated cells in the presence of proteins derived from lymphocytes.

Protein synthesis of nylon-wool adherent (B) and non-adherent (T) human tonsillar lymphocytes was examined in the presence of three protein fractions precipitated with ammonium sulfate from the medium of the above lymphocytes incubated in vitro for 4 hours. One fraction precipitated with ammonium sulfate at 0--30% saturation was found to inhibit amino acid incorporation into proteins mainly in unseparated cells while the fraction precipitated at 30--70% saturation decreased the rate of protein synthesis in B cells. The fraction precipitated at 70--100% saturation inhibited protein synthesis slightly in T lymphocytes. Protein synthesis in B and T as well as in unseparated cells was also examined in the presence of proteins obtained from the medium of B and T lymphocytes. It is assumed that proteins released by lymphocytes without mitogenic activation in vitro are involved in the mediation of lymphocyte interactions and may be related to lymphokines synthesized and released by activated cells.

B-Lymphocytes↗