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101.
Craig R. Halberstadt Bernhard O. Palsson A. Rees Midgley Rane L. Curl 《Biotechnology and Bioprocess Engineering》2002,7(3):163-170
This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of
medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained
in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence
time distribution similar to a continuous stirred tank reactor (CSTR). However, due to the arrangement of the medium lines
in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from
short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence
times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided
into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides
nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes
of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed
from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases
as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter.
As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular
metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation
of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects
on cellular metabolism. 相似文献
102.
A variety of processes have been proposed for mammalian cell culture in the commercial production of useful substances (e.g., monoclonal antibodies, therapeutic and diagnostics proteins). Among them, the perfusion culture of suspended non-immobilized cells is the most advantageous. Perfusion culture can be classified by the separation process of suspended cells from the culture mixture into three types, namely filtration, gravitational settling and centrifugation. From a commercial point of view, the present situation and technical problems of suspended-cell perfusion culture will be reviewed based on the three types, The recent development of perfusion culture has been carried out mainly on the filtration separation process, but the centrifugation process seems to have a promising future because of operation stability and scale-up feasibility. The reasons will be explained in details. 相似文献
103.
Substance P has been implicated as a neuronal mediator of inflammation in various inflammatory conditions. However, the exact role played by substance P in inflammatory bowel diseases or in experimental colonic vasculitis has not been clearly understood. In this study, we examined the effect of close superior mesenteric artery injection of substance P under prevailing inflammatory conditions induced by intravenous human albumin antialbumin immune complex followed by intracolonic perfusion of 2.5% formaldehyde in rats or intracolonic perfusion of 5% alcohol alone. The immune complex- and formaldehyde-treated rats showed severe microvascular changes such as microvascular plugging by red blood cells, endothelial breakage and extravasation of plasma proteins and red blood cells. The bolus injection of 10−8 M substance P reduced extravasation of Evans blue dye by 50% and the tissue wet to dry ratio by 20% in immune complex- and formaldehyde-perfused rats. Myeloperoxidase activity was not changed. Substance P also significantly inhibited (44%) the extravasation in alcohol-perfused rats. Pretreatment of immune complex- and formaldehyde-treated rats with substance P antagonist reversed the effect of substance P. These findings suggest that the most immediate effect of substance P may be vasodilation and clearing of vascular plugs induced by immune complex and formaldehyde. This effect of substance P differs from its chronic effect, which causes vasodilation and extravasation. 相似文献
104.
Alexander Nikolay Joris de Grooth Yvonne Genzel Jeffery A. Wood Udo Reichl 《Biotechnology and bioengineering》2020,117(10):3040-3052
The use of bioreactors coupled to membrane-based perfusion systems enables very high cell and product concentrations in vaccine and viral vector manufacturing. Many virus particles, however, are not stable and either lose their infectivity or physically degrade resulting in significant product losses if not harvested continuously. Even hollow fiber membranes with a nominal pore size of 0.2 µm can retain much smaller virions within a bioreactor. Here, we report on a systematic study to characterize structural and physicochemical membrane properties with respect to filter fouling and harvesting of yellow fever virus (YFV; ~50 nm). In tangential flow filtration perfusion experiments, we observed that YFV retention was only marginally determined by nominal but by effective pore sizes depending on filter fouling. Evaluation of scanning electron microscope images indicated that filter fouling can be reduced significantly by choosing membranes with (i) a flat inner surface (low boundary layer thickness), (ii) a smooth material structure (reduced deposition), (iii) a high porosity (high transmembrane flux), (iv) a distinct pore size distribution (well-defined pore selectivity), and (v) an increased fiber wall thickness (larger effective surface area). Lowest filter fouling was observed with polysulfone (PS) membranes. While the use of a small-pore PS membrane (0.08 µm) allowed to fully retain YFV within the bioreactor, continuous product harvesting was achieved with the large-pore PS membrane (0.34 µm). Due to the low protein rejection of the latter, this membrane type could also be of interest for other applications, that is, recombinant protein production in perfusion cultures. 相似文献
105.
A 15.6 frames per second 1‐megapixel multiple exposure laser speckle contrast imaging setup
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Martin Hultman Ingemar Fredriksson Marcus Larsson Atila Alvandpour Tomas Strömberg 《Journal of biophotonics》2018,11(2)
A multiple exposure laser speckle contrast imaging (MELSCI) setup for visualizing blood perfusion was developed using a field programmable gate array (FPGA), connected to a 1000 frames per second (fps) 1‐megapixel camera sensor. Multiple exposure time images at 1, 2, 4, 8, 16, 32 and 64 milliseconds were calculated by cumulative summation of 64 consecutive snapshot images. The local contrast was calculated for all exposure times using regions of 4 × 4 pixels. Averaging of multiple contrast images from the 64‐millisecond acquisition was done to improve the signal‐to‐noise ratio. The results show that with an effective implementation of the algorithm on an FPGA, contrast images at all exposure times can be calculated in only 28 milliseconds. The algorithm was applied to data recorded during a 5 minutes finger occlusion. Expected contrast changes were found during occlusion and the following hyperemia in the occluded finger, while unprovoked fingers showed constant contrast during the experiment. The developed setup is capable of massive data processing on an FPGA that enables processing of MELSCI data in 15.6 fps (1000/64 milliseconds). It also leads to improved frame rates, enhanced image quality and enables the calculation of improved microcirculatory perfusion estimates compared to single exposure time systems. 相似文献
106.
G. Bonardel C. Barrau M. Soussan E. D’Estanque B. Erra C. Etard N. Fayard M.O. Habert S. Hapdey E. Hindie F. Hyafil C. Labriolle Vaylet O. Lairez P.Y. Leroux P. Olivier M. Queneau E. Rust R. Seban M. Wartski 《Médecine Nucléaire》2018,42(5):354-358
The examples and clinical cases presented in this section are not intended to be considered as absolute models in terms of image quality or device parameter settings. They must initiate an individual analysis according to CT parameters and image quality. Nevertheless, they present practically different CT levels, which can be used according to the clinical context and the type of device. 相似文献
107.
G. Bonardel C. Barrau M. Soussan E. D’Estanque B. Erra C. Etard N. Fayard M.O. Habert S. Hapdey E. Hindie F. Hyafil C. Labriolle Vaylet O. Lairez P.Y. Leroux P. Olivier M. Queneau E. Rust R. Seban M. Wartski 《Médecine Nucléaire》2018,42(5):370-378
The examples and clinical cases presented in this section are not intended to be considered as absolute models in terms of image quality or device parameter settings. They must initiate an individual analysis according to CT parameters and image quality. Nevertheless, they present practically different CT levels which can be used according to the clinical context and the type of device. 相似文献
108.
K. Akiyama T.L. Gluckman A. Terhakopian P.M. Jinadasa S. Narayan S. Singaswamy B. Massey III R.J. Bing 《Tissue & cell》1997,29(6):733-743
We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions. 相似文献
109.
缺氧条件下冻伤对大鼠微循环血液灌流量的影响 总被引:1,自引:1,他引:0
本文采用体重200±20g健康雄性Wistar大鼠,随机分为平原冻伤(FN)组、急性缺氧冻伤(FAH)组和缺氧习服缺氧冻伤(FHAC)组,实验观察了大鼠右后肢重度冻伤前后各组大鼠双后肢皮肤微循环灌流量的改变。结果表明,平原冻伤使大鼠双后肢微循环灌流量明显减少,提示局部重度冻伤对微循环的影响不只局限于冻区也涉及到对侧肢体。冷冻前FAH组大鼠微循环灌流量已明显低于FN组,表明急性缺氧时血容量进行代偿性的再分配,使微循环灌流量减少;FAH组大鼠冻后双后肢微循环灌流量的改变结果提示急性缺氧可加重其冻伤对微循环的损伤程度。FHAC组大鼠冻前微循环灌流量非常明显地低于正常对照,也明显低于急性缺氧对照,表明缺氧习服可造成微循环障碍;FHAC组大鼠冻肢微循环灌流量非常明显地低于FN组,提示缺氧习服加重高原冻伤引起的微循环障碍。 相似文献
110.
Karl Schumacher Raimund Strehl Sabine Kloth Michel Tauc Will W. Minuth 《In vitro cellular & developmental biology. Animal》1999,35(8):465-471
Summary During kidney development the embryonic ampullar collecting duct (CD) epithelium changes its function. The capability for
nephron induction is lost and the epithelium develops into a heterogeneously composed epithelium consisting of principal and
intercalated cells. Part of this development can be mimicked under in vitro conditions, when embryonic collecting duct epithelia
are isolated from neonatal rabbit kidneys and kept under perfusion culture. The differentiation pattern is quite different
when the embryonic collecting duct epithelia are cultured in standard Iscove’s modified Dulbecco’s medium as compared to medium
supplemented with additional NaCl. Thus, the differentiation behavior of embryonic CD epithelia is unexpectedly sensitive.
To obtain more information about how much influence the medium has on cell differentiation, we tested medium 199, basal medium
Eagle, Williams’ medium E, McCoys 5A medium, and Dulbecco’s modified Eagle medium under serum-free conditions. The experiments
show that in general, all of the tested media are suitable for culturing embryonic collecting duct epithelia. According to
morphological criteria, there is no difference in morphological epithelial cell preservation. The immunohistochemical data
reveal two groups of expressed antigens. Constitutively expressed antigens such as cytokeratin 19, PCD 9, Na/K ATPase, and laminin are present in all cells of the epithelia independent of the culture media used. In contrast,
a group of antigens detected by mab 703, mab 503, and PNA is found only in individual series. Thus, each culture medium produces
epithelia with a very specific cell differentiation pattern. 相似文献