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1.
The uptake of [3H]norepinephrine ([3H]NE) was studied in dissociated brain cell cultures prepared from 8-day-old chick embryos using the whole brain (minus optic lobes). Uptake of [3H]NE, 5×10–9 M, 10 min incubation, in freshly dissociated noncultured embryonic chick brain cells, was detected in 6-day-old embryos; it was temperature and drug (cocaine, metanephrine) sensitive and increased with brain development. In cultured cells, which were assayed at various days in culture, the increase in [3H]NE accumulation per culture was less than that seen in freshly dissociated noncultured embryonic cells. When [3H]NE uptake was expressed per mg protein, a decrease with days in culture was observed, reflecting perhaps a dilution of growth or proliferation of cells not accumulating NE. Metanephrine, 5×10–6 M, an inhibitor of extraneuronal uptake, inhibited [3H]NE in 5-day-old cultures whereas desmethylimipramine, an inhibitor of neuronal uptake, inhibited [3H]NE uptake in 15- and 20-day-old cultures. Cocaine, another neuronal inhibitor, inhibited [3H]NE at 10 and 15 days only. We interpret these findings to suggest that during early growth in culture most neuroblasts accumulate NE nonspecifically and, as neuronal maturation proceeds, NE accumulation becomes specific. 相似文献
2.
Brain extracts from 8-day-old chick embryos have been shown to influence morphological development of dissociated brain cells from 7-day-old chick embryos in culture. Stimulatory, effects on size of the neuronal somas and on growth of long processes were observed by adding the cytosol of the brain extract or the dialysate of the cytosol. These morphological changes parallel modifications of various enzyme activities according to the age of the cultures. Adenyl cyclase, (Na+, K+)- and Mg2+-ATPase, 5-nucleotidase, choline acetyltransferase, and acetylcholinesterase activities were studied between 5 and 14 days of culture. Adenyl cyclase activity was strongly stimulated at 8 days by both extracts. (Na+, K+)-and Mg2+-ATPase activities were stimulated in 8-day-old cultures only by the dialysate. 5-Nucleotidase activity was stimulated in 8-day-old cultures by the dialysate and in 11-day-old cultures by both extracts. Choline acetyltransferase activity was stimulated by the cytosol in 8-day-old cultures and by the dialysate in 11-day-old cultures. The total acetylcholinesterase activity was higher in 8-, 11-, and 14-day-old cultures treated with the cytosol. When the cells were treated with the dialysate, the activity was only higher in 14-day-old cultures. We also found that following the addition of brain extracts, the specific activity of the enzymes we studied was enhanced and became close to the values found in vivo during embryogenesis. Thus in parallel to the morphological modifications observed in nerve cell cultures treated by embryo brain extracts, biochemical variations especially involved in synaptogenesis and membrane development could be measured. 相似文献
3.
Hélène Perreten Lambert Manuel Zenger Guillaume Azarias Jean-Yves Chatton Pierre J. Magistretti Sylvain Lengacher 《The Journal of biological chemistry》2014,289(45):31014-31028
Brain activity is energetically costly and requires a steady and highly regulated flow of energy equivalents between neural cells. It is believed that a substantial share of cerebral glucose, the major source of energy of the brain, will preferentially be metabolized in astrocytes via aerobic glycolysis. The aim of this study was to evaluate whether uncoupling proteins (UCPs), located in the inner membrane of mitochondria, play a role in setting up the metabolic response pattern of astrocytes. UCPs are believed to mediate the transmembrane transfer of protons, resulting in the uncoupling of oxidative phosphorylation from ATP production. UCPs are therefore potentially important regulators of energy fluxes. The main UCP isoforms expressed in the brain are UCP2, UCP4, and UCP5. We examined in particular the role of UCP4 in neuron-astrocyte metabolic coupling and measured a range of functional metabolic parameters including mitochondrial electrical potential and pH, reactive oxygen species production, NAD/NADH ratio, ATP/ADP ratio, CO2 and lactate production, and oxygen consumption rate. In brief, we found that UCP4 regulates the intramitochondrial pH of astrocytes, which acidifies as a consequence of glutamate uptake, with the main consequence of reducing efficiency of mitochondrial ATP production. The diminished ATP production is effectively compensated by enhancement of glycolysis. This nonoxidative production of energy is not associated with deleterious H2O2 production. We show that astrocytes expressing more UCP4 produced more lactate, which is used as an energy source by neurons, and had the ability to enhance neuronal survival. 相似文献
4.
Roxana Mamani Anccasi Isis Moraes Ornelas Marcelo Cossenza Pedro Muanis Persechini Ana Lucia Marques Ventura 《Purinergic signalling》2013,9(1):15-29
Previous data suggest that nucleotides are important mitogens in the developing retina. Here, the effect of ATP on the death of cultured chick embryo retina cells was investigated. In cultures obtained from retinas of 7-day-old chick embryos (E7) that were cultivated for 2 days (E7C2), both ATP and BzATP induced a ~30 % decrease in cell viability that was time- and dose-dependent and that could be blocked by 0.2 mM oxidized ATP or 0.3 μM KN-62. An increase in cleaved caspase-3 levels and in the number of TUNEL-positive cells was observed when cultures were incubated with 3 mM ATP and immunolabeling for cleaved-caspase 3 was observed over neurons but not over glial cells. ATP-dependent cell death was developmentally regulated, the maximal levels being detected by E7C2-3. Nucleotides were able to increase neuronal ethidium bromide and sulforhodamine B uptake in mixed and purified neuronal cultures, an effect that was blocked by the antagonists Brilliant Blue G and oxidized ATP. In contrast, nucleotide-induced cell death was observed only in mixed cultures, but not in purified cultures of neurons or glia. ATP-induced neuronal death was blocked by the glutamatergic antagonists MK801 and DNQX and activation of P2X7 receptors by ATP decreased the uptake of [3H]-d-aspartate by cultured glial cells with a concomitant accumulation of it in the extracellular medium. These results suggest that ATP induces apoptosis of chick embryo retinal neurons in culture through activation of P2X7 and glutamate ionotropic receptors. Involvement of a P2X7 receptor-mediated inhibition of the glial uptake of glutamate is suggested. 相似文献
5.
The uptake of calcium was examined in primary cultures of pure neurons and of glial cells from dissociated hemispheres of chick embryo brain. Neuronal cultures took up calcium at a rate of 2.0 nmol per min per mg cell protein at medium concentrations of 1.2 mM-Ca2+ and 5.4 mM-K+. The rate of calcium entry into neurons was increased 2.7-fold by elevating medium potassium to 60 MM. The effect of high external potassium was to increase the Vmax value for calcium transport from 5.5 to 13 nmol per min per mg; the Michaelis constant for calcium, 1.2 mM, was unchanged. The potassium-dependent component of calcium entry into the neuronal cultures was eliminated by addition of 0.1 mM-D-600 (a verapamil derivative) or by 1 mM-CoCl2, but 0.5 μM-tet-rodotoxin had no significant effect. When choline replaced potassium in uptake medium no change in calcium transport was detected in neurons, nor was the entry of calcium increased when choline replaced sodium. Glial cultures took up calcium at 20% of the basal rate for neuronal cultures on a weight-of-protein basis. Uptake was not increased by potassium; during depolarization by potassium the calcium transport activity of glia was less than 10% that of neurons. It was concluded that cultured neurons contain a depolarization-sensitive, calcium-specific channel. A similar calcium transport activity was not detected in cultured glial cells. 相似文献
6.
Whole cells ofTreponema pallidum consumed O2 with lactate in a glucose-depleted medium.d(–) Lactate caused marked stimulation of O2 uptake at a rate similar to that with glucose, whereasl(+) lactate resulted in no increase over the reduced rate observed upon glucose depletion. Lactate oxidation was specific for -hydroxy straight-chain acids of 3,4, and 5 carbons. O2 uptake during lactate oxidation proceeded independently of pyruvate oxidation and required NAD. The product of lactate oxidation was pyruvate.d(–) Lactate-stimulate O2 uptake was sensitive to chlorpromazine and resistant to amytal and cyanide. Glucose did not inhibit the oxidation of lactate as shown by the additive effect of both substrates on O2 uptake. Oxidation of glucose, but not lactate, provided energy necesary for motilibty or maintenance of virulence. A mixture of lactate isomers was formed from glucose with thel(+) isomer concentration remaining constant and thed(–) isomer concentration varying inversely with dissolved O2 concentration. The function of lactate as an oxidizable substrate is apparently quite distinct from that of glucose. 相似文献
7.
Protoveratrine-(5 M) stimulated aerobic glycolysis of incubated rat brain cortex slices that accompanies the enhanced neuronal influx of Na+ is blocked by tetrodotoxin (3 M) and the local anesthetics, cocaine (0.1 mM) and lidocaine (0.5 mM). On the other hand, high [K+]-stimulated aerobic glycolysis that accompanies the acetylcholine-sensitive enhanced glial uptakes of Na+ and water is unaffected by acetylcholine (2 mM). Experiments done under a variety of metabolic conditions show that there exists a better correlation between diminished ATP content of the tissue and enhanced aerobic glycolysis than between tissue ATP and the ATP-dependent synthesis of glutamine. Whereas malonate (2 mM) and amino oxyacetate (5 mM) suppress ATP content and O2 uptake, stimulate lactate formation, but have little effect on glutamine levels, fluoroacetate (3 mM) suppresses glutamine synthesis in glia, presumably by suppressing the operation of the citric acid cycle, with little effect on ATP content, O2 uptake, and lactate formation. Exogenous citrate (5 mM), which may be transported and metabolized in glia but not in neurons, inhibits lactate formation by cell free acetone-dried powder extracts of brain cortex but not by brain cortex slices. These results suggest that the neuron is the major site of stimulated aerobic glycolysis in the brain, and that under our experimental conditions glycolysis in glia is under lesser stringent metabolic control than that in the neuron. Stimulation of aerobic glycolysis by protoveratrine occurs due to diminution of the energy charge of the neuron as a result of stimulation of the sodium pump following tetrodotoxin-sensitive influx of Na+; stimulation by high [K+, NH4
+, or Ca2+ deprivation occurs partly by direct stimulation of key enzymes of glycolysis and partly by a fall in the tissue ATP concentration. 相似文献
8.
Lüdemann I Pörtner R Schaefer C Schick K Srámkova K Reher K Neumaier M Franěk F Märkl H 《Cytotechnology》1995,19(2):111-124
A murine hybridoma cell line producing a monoclonal antibody against penicillin-G-amidase and a murine transfectoma cell line secreting a monovalent chimeric human/mouse Fab-antibody fragment were cultivated in three different media (serum-containing, low protein serum-free, and iron-rich protein-free) in flask cultures, stirred reactors and a fixed bed reactor. In static batch cultures in flasks both cell lines showed similar good growth in all three media.In suspension in a stirred reactor, the hybridoma cell line could be cultivated satisfactory only in serum-containing medium. In low protein serum-free medium, Pluronic F68 had to be added to protect the hybridoma cells against shear stress. But even with this supplement only batch, not chemostat mode was possible. In iron-rich protein-free medium the hybridoma cells grew also in continuous chemostat mode, but the stability of the culture was low. The transfectoma cell line did not grow in stirred reactors in any of the three media.Good results with both cell lines were obtained in fixed bed experiments, where the cells were immobilized in macroporous Siran®-carriers. The media, which were optimized in flask cultures, could be used without any further adaptation in the fixed bed reactor. Immobilization improved the stability and reliability of cultures of non-adherent animal cells in serum-free media tremendously compared to suspension cultures in stirred reactors. The volume-specific glucose uptake rate, an, indicator of the activity of the immobilized cells, was similar in all three media. Deviations in the metabolism of immobilized and suspended cells seem to be mainly due to low oxygen concentrations within the macroporous carriers, where the cells are supplied with oxygen only by diffusion.List of symbols c
substrate or product concentration mmol l–1
- c0
substrate or product concentration in the feed mmol l–1
- cGlc
glucose concentration mmol l–1
- cGln
glutamine concentration mmol l–1
- cAmm
ammonia concentration mmol l–1
- cLac
lactate concentration mmol l–1
- cFAB
concentration of Fab# 10 antibody fragment g l–1
- cMAb
monoclonal antibody concentration mg l–1
- D
dilution rate d–1
- q
cell-specific substrate uptake or metabolite production rate mmol cell–1 h–1
- qGlc
cell-specific glucose uptake rate mmol cell–1 h–1
- qGln
cell-specific glutamine uptake rate mmol cell–1 h–1
- qMAb
cell-specific MAb production rate mg cell–1 h–1
- q*
volume-specific substrate uptake or metabolite production rate mmol l–1 h–1
- q*FB
volume-specific substrate uptake or metabolite production rate related to the fixed bed volume mmol lFB
–1 h–1
- q*FB,Glc
volume-specific glucose uptake rate related to the fixed bed volume mmol lFB
–1 h–1
- q*FB,Gln
volume-specific glutamine uptake rate related to the fixed volume mmol lFB
–1 h–1
- q*FB,MAb
volume-specific MAb production rate related to the fixed volume mg lFB
–1 h–1
- q*FB,02
volume-specific oxygen uptake rate related to the fixed bed volume mmol lFB
–1 h–1
- t
time h
- U
superficial flow velocity mm s–1
- V
medium volume in the conditioning vessel of the fixed bed reactor l
- VFB
volume of the fixed bed l
- xv
viable cell concentration cells ml–1
- yAmm,Gln
yield of Ammonia from glutamine
- yLac,Glc
yield of lactate from glucose
-
specific growth rate h–1
- d
specific death rate h–1 相似文献
9.
Rapid changes in oxidative metabolism as a consequence of elicitor treatment of suspension-cultured cells of French bean (Phaseolus vulgaris L.) 总被引:1,自引:0,他引:1
Duncan Robertson Dewi R. Davies Chris Gerrish Steven C. Jupe G. Paul Bolwell 《Plant molecular biology》1995,27(1):59-67
Stressed plant cells often show increased oxygen uptake which can manifest itself in the transient production of active oxygen species, the oxidative burst. There is a lack of information on the redox status of cells during the early stages of biotic stress. In this paper we measure oxygen uptake and the levels of redox intermediates NAD/NADH and ATP and show the transient induction of the marker enzyme for redox stress, alcohol dehydrogenase. Rapid changes in the redox potential of elicitor-treated suspension cultures of French bean cells indicate that, paradoxically, during the period of maximum oxygen uptake the levels of ATP and the NADH/NAD ratio fall in a way that indicates the occurrence of stress in oxidative metabolism. This period coincides with the maximum production of active oxygen species particularly H2O2. The cells recover and start producing ATP immediately upon the cessation of H2O2 production. This indicates that the increased O2 uptake is primarily incorporated into active O2 species. A second consequence of these changes is probably a transient compromising of the respiratory status of the cells as indicated in expression of alcohol dehydrogenase. Elicitor-induced bean ADH was purified to homogeneity and the Mr 40 000 polypeptide was subjected to amino acid sequencing. 15% of the whole protein was sequenced from three peptides and was found to have nearly 100% sequence similarity to the amino acid sequence for pea ADH1 (PSADH1). The cDNA coding for the pea enzyme was used to demonstrate the transient induction of ADH mRNA in elicitor-treated bean cells. Enzyme activity levels also increased transiently subsequently. Increased oxygen uptake has previously been thought to be associated with provision of energy for the changes in biosynthesis that occur rapidly after perception of the stress signal. However the present work shows that this rapid increase in oxygen uptake as a consequence of elicitor action is not wholly associated with respiration. 相似文献
10.
Wibke Wagner Paul Ajuh Johannes Löwer Silja Wessler 《Cell communication and signaling : CCS》2010,8(1):1-9
Following cultivation of distinct mesenchymal stem cell (MSC) populations derived from human umbilical cord under hypoxic conditions (between 1.5% to 5% oxygen (O2)) revealed a 2- to 3-fold reduced oxygen consumption rate as compared to the same cultures at normoxic oxygen levels (21% O2). A simultaneous measurement of dissolved oxygen within the culture media from 4 different MSC donors ranged from 15 μmol/L at 1.5% O2 to 196 μmol/L at normoxic 21% O2. The proliferative capacity of the different hypoxic MSC populations was elevated as compared to the normoxic culture. This effect was paralleled by a significantly reduced cell damage or cell death under hypoxic conditions as evaluated by the cellular release of LDH whereby the measurement of caspase3/7 activity revealed little if any differences in apoptotic cell death between the various cultures. The MSC culture under hypoxic conditions was associated with the induction of hypoxia-inducing factor-alpha (HIF-1α) and an elevated expression of energy metabolism-associated genes including GLUT-1, LDH and PDK1. Concomitantly, a significantly enhanced glucose consumption and a corresponding lactate production could be observed in the hypoxic MSC cultures suggesting an altered metabolism of these human stem cells within the hypoxic environment. 相似文献
11.
The efficiency of (Na+ + K+)-ATPase (i.e. the amount of K+ pumped per ATP hydrolyzed) in intact tumorigenic cells was estimated in this study. This was accomplished by simultaneously measuring the rate of ouabain-sensitive K+ uptake and oxygen consumption in tumorigenic cell suspensions during the reintroduction of K+ to K+-depleted cells. The ATP turnover was then estimated by assuming 5.6–6 ATP/O2 as the stoichiometry of NADH-linked respiration in these cells. In the three cell lines tested (hamster and chick embryo cells transformed with Rous sarcoma virus and Ehrlich ascites cells), the K+/ATP ratio was approximately 2, the same value as that found in normal tissues. Furthermore, only 20% of the total ATP production of these cells was used by (Na+ + K+)-ATPase. 相似文献
12.
Primary cultures of glial cells prepared from brains of newborn rats were grown for periods of 1–5 weeks. After a proliferative phase of between 2 and 3 weeks, the cultures were maintained in stationary phase, during which a significant increase of oxygen consumption and of the activities of lactate dehydrogenase, succinate dehydrogenase, and mitochondrial glycerolphosphate dehydrogenase could be observed. Furthermore, qualitative changes in the lactate dehydrogenase isoenzyme pattern were found with time, characterized by a shift toward an enhanced synthesis of H subunits. A similar development was found in comparing the LDH isoenzyme pattern in the brain of 15-day-old rat embryo with those of newborn and adult rat brains. It is suggested that some aspects of maturation of glial cells in culture are comparable to those occurring in whole brain in vivo, namely a shift towards an enhanced aerobic metabolism. 相似文献
13.
U. Karsten A. Kössler E. Janiszewski A. Wollenberger 《In vitro cellular & developmental biology. Plant》1973,9(3):139-146
Summary Primary cell cultures from neonatal rat ventricles were continuously exposed for 7 days in a modified roller apparatus to
defined pericellular oxygen tension varying from 0.6 to 600 mm Hg. 5-Fluorodeoxyuridine was added to the medium to prevent
over-growth of muscle cells by nonmuscle cells. A pericellular pO2 of 600 mm Hg was lethal. The range of about 15 to 150 mm Hg was favorable, as indicated by increases in total and muscle-characteristic
proteins. Between the 2nd and 8th day of cultivation at a pO2 of 38 mm Hg, myosin content per cell increased 3.2-fold and creatine kinase activity 2.5-fold. At 0.6 mm Hg, myosin content
increased only 1.3-fold and there was no increase in creatine kinase activity. The rate of myosin synthesis was diminished
at this low pO2. ATP level and beating rate at 0.6 mm Hg did not differ from values at 38 mm Hg. The isoenzyme pattern of lactate dehydrogenase
remained unchanged during cultivation at 38 mm Hg, whereas at 0.6 mm Hg it shifted towards an M-type pattern. These experiments
suggest that neonatal rat heart cells maintained in vitro can adapt themselves to low oxygen tensions. 相似文献
14.
FSH , but not LH, increased the O2 uptake of isolated granulosa cells from 23 day old rats previously treated with DES or with DES and FSH. Dose response studies showed that the cells were most sensitive to FSH when the cellular binding of FSH was highest. LH increased the O2 uptake of granulosa cells of untreated 30 day old rats. DES treatment inhibited the LH induced rise in O2 uptake when the rats were implanted with DES capsules unless FSH was injected to induce LH receptors. Addition of dbcAMP increased O2 uptake of granulosa cells from 30 day old rats at concentrations 10X lower than those required to stimulate O2 uptake in cells from 23 day old rats treated with DES alone.FSH increased lactate formation in the absence of added substrates but did not do so when glucose was added to the media. In contrast, LH greatly increased lactate formation with added glucose. Dose response studies showed that less than 0.6 ug/ml LH S21 was effective in increasing lactate above control levels. These data suggest that FSH affects aerobic pathways while LH affects anaerobic pathways in the process of the differentiation of granulosa cells toward luteal cells.It is well known that FSH and LH interact with their target cells in the ovary by binding to specific receptors and that FSH stimulates LH-receptor production (1). Receptor binding by either hormone activates adenylate cyclase (2) raising cyclic adenosine monosphosphate (cAMP) levels (3) and increasing protein kinase activity (4). Such changes probably trigger changes in the major metabolic pathways that support follicular development because cells of corpora lutea have glycogen (5) which is not present in follicular granulosa cells (6–9). Several studies suggest that FSH and LH may regulate metabolic processes in the ovary. LH increases lactate in whole prepuberal ovaries (10,11,12) and also increases the uptake of glucose (13). FSH increases oxygen uptake in chick ovaries (14), rat ovaries (15) and prairie dog ovaries (16). However, only one study has been done using isolated ovarian cells. Hamberger (17) has reported that FSH increased the oxygen uptake of thecal cells of immature rats while LH increased the oxygen uptake of granulosa cells. Since granulosa cells from immature rats are reported to have FSH receptors while theca cells have LH receptors the effects of these hormones appear unclear.The present studies were undertaken to more accurately characterize the actions of FSH, LH, and dibutyryl cAMP (dbcAMP) on the oxygen uptake of isolated granulosa cells and remaining tissues of immature ovaries and to determine the effects of FSH and LH on the production of lactate by granulosa cells. 相似文献
15.
Evaluation of monitoring approaches and effects of culture conditions on recombinant protein production in baculovirus-infected insect cells 总被引:2,自引:0,他引:2
The baculovirus infection process ofSpodoptera frugiperda (Sf9) insect cells in oxygen-controlled bioreactors in serum-free medium was investigated using a recombinantAutographa californica (AcNPV) virus expressing -galactosidase enzyme as a model system. A variety of monitoring techniques including trypan blue exclusion, fluorescent dye staining, oxygen uptake rate (OUR) measurements, and glucose consumption were applied to infected cells to determine the best way of evaluating cell integrity and assessing the course of baculovirus infection. The metabolism of newly-infected cells increased 90% during the first 24 hours, but as infection proceeded, and cells gradually succumbed to the baculovirus infection, the cytopathic effect of the baculovirus on the cells became evident. Oxygen and glucose uptake rate measurements appeared to more accurately assess the condition of infected cells than conventional trypan blue staining, which tended to overestimate cell viability in the mid stages of infection. The optimal harvest time varied, depending on which technique — SDS-PAGE, chromogenic (ONPG) or fluorometric (C12FDG) — was used to monitor -galactosidase production. Specific -galactosidase production was found to be insensitive to a wide range of culture dissolved oxygen tensions, whereas resuspending cells in fresh medium prior to infection increased volumetric productivity approximately two-fold (800,000 units -galactosidase/ml) compared to cultures infected in batch mode and allowed successful infections to occur at higher cell densities.Abbreviations ONPG
ortho-phenyl 2--D-galactopyranoside
- OUR
oxygen uptake rate (-mol O2/liter/hour)
- qglucose
specific glucose uptake rate (mg glucose/106cell/hour)
- qglutamine
specific glutamine uptake rate (mg glutamine/106cell/hour)
- qO2
specific oxygen uptake rate (-mol O2/106cell/hour)
- MOI
virus multiplicity of infection (viral plaque forming units/cell) 相似文献
16.
On-line estimation of viable cells in a hybridoma culture at various DO levels using ATP balancing and redox potential measurement 总被引:1,自引:0,他引:1
Two on-line methods for the estimation of viable cell number in hybridoma cultivation were investigated. One used an empirical correlation between redox potential and animal cell density. The other was based on an ATP balance with ATP steady-state assumption. Oxygen uptake rate measurement provided the amount of ATP which was produced by oxidation of NADH. Oxygen uptake rate was measured either by stationary liquid phase balance with surface aeration or by gas balance during bubble aeration with headspace flushing with an inert gas. The amount of ATP produced through the glycolysis was estimated based on the amount of lactate produced. In cultures, in which pH was controlled via manipulation of the gas phase composition, the flow of CO(2) was linearly correlated with the lactate concentration. At constant dissolved oxygen levels, the viable cell density was proportional to the estimated ATP production rate, during exponential growth and during later phases. The estimated specific ATP production rate, however, varied from 2.2 pmol cell(-1) h(-1) at 10% air saturation to 4.5 pmol cell(-1) h(-1) at 100% air saturation. Specific rates of glutamine, glucose, and lactate followed the shape of the specific ATP production rate, whereas the specific oxygen uptake rate was minimal at around 50% air saturation. (c) 1996 John Wiley & Sons, Inc. 相似文献
17.
Oxygen Dependence of Glucose and Acetylcholine Metabolism in Slices and Synaptosomes from Rat Brain 总被引:12,自引:10,他引:2
Abstract: Previous studies have shown that a reduction in the O2 tension of the blood from 120 torr to 57 torr (hypoxic hypoxia) decreases brain acetylcholine (ACh) synthesis. To determine if this decrease is due to a direct impairment of ACh metabolism or to an indirect effect mediated by other neurotransmitter systems, we studied ACh formation in rat brain slices and synaptosomes. At O2 tensions ranging from 760 to less than 1 torr, 14CO2 production and [14C]ACh synthesis from [U-14C]glucose, the levels of lactate and ATP, and the ATP/ADP ratio were determined. In slices, the first decreases were observed in the rate of 14CO2 production and [14C]ACh synthesis at an O2 tension of 152 torr. The ATP level started to decline at 53–38 torr, and a reduction in the ATP/ADP ratio was first found at and below 19 torr. Lactate formation was maximally stimulated at 38–19 torr. Synaptosomes responded differently than brain slices to reduced O2 tensions. In synaptosomes, 14CO2 production and [14C]ACh synthesis from [U-14C]glucose, the levels of lactate and ATP, and the ATP/ADP ratio were unaltered if a minimum O2 tension of 19 torr was maintained. Despite the difference in sensitivities to decreases in O2 levels, there is a curvilinear relationship between [U-14C]glucose decarboxylation and [14C]ACh synthesis at various O2 tensions for both tissue preparations with a high coefficient of determination (R2= 0.970). The difference in the metabolic sensitivity of slices and synaptosomes to a reduced O2 level may be explained by the greater distance O2 must diffuse in slices. The results are discussed in comparison with hypoxia in vivo. 相似文献
18.
The ample supply of O2 proved to be of great importance for berberine production in cell suspension culture ofThalictrum minus, as the specific O2 consumption rate of berberine-producing cells was twice as high as that of non-producing cells. Furthermore, berberine yield increased with increases in the volumetric O2 transfer coefficient (KLa). Estimation of the optimum conditions of oxygen supply in suspension cultures and immobilized cells according to a known theoretical model assuming O2 uptake by cells to be a zero-order reaction was in good agreement with the experimental data. The O2 supply to immobilized cells could be improved by reducing the cell density and radius of the bead. 相似文献
19.
The cytotoxic effect of high, as well as low, oxygen tension of proliferation and metabolism of Low line cells in culture is reversible even after several days of exposure provided the cells are returned to 95% air + 5% CO2 environment. This suggests that the activity of certain mechanisms within the cells may have been altered or in other ways inhibited by the abnormal environments but are quite rapidly regenerated once the adverse condition is removed. The cells tolerate a low O2 exposure for at least 20 days while continuous exposure to high O2 atmosphere results in degeneration and death after 7–10 days. Both glucose utilization and lactic acid production are elevated in cultures exposed to either low or high O2 tensions, although they are markedly higher in the latter condition. When cell so exposed are returned to an air + 5% CO2 atmosphere, rate of glucose uptake and lactic acid formation soon approaches that found in control cultures. 相似文献
20.
Dialysis cultures with immobilized hybridoma cells for effective production of monoclonal antibodies
An industrial scale reactor concept for continuous cultivation of immobilized animal cells (e.g. hybridoma cells) in a radial-flow fixed bed is presented, where low molecular weight metabolites are removed via dialysis membrane and high molecular products (e.g. monoclonal antibodies) are enriched. In a new nutrient-split feeding strategy concentrated medium is fed directly to the fixed bed unit, whereas a buffer solution is used as dialysis fluid. This feeding strategy was investigated in a laboratory scale reactor with hybridoma cells for production of monoclonal antibodies. A steady state monoclonal antibody concentration of 478 mg l-1 was reached, appr. 15 times more compared to the concentration reached in chemostat cultures with suspended cells. Glucose and glutamine were used up to 98%. The experiments were described successfully with a kinetic model for immobilized growing cells. Conclusions were drawn for scale-up and design of the large scale system.Abbreviations: cGlc – glucose concentration, mmol l-1; cGln – glutamine concentration, mmol l-1; cAmm – ammonia concentration, mmol l-1; cLac – lactate concentration, mmol l-1; cMAb – MAb concentration, mg l-1; D – dilution rate, d-1; Di – dilution rate in the inner chamber of the membrane dialysis reactor, d-1; D0 – dilution rate in the outer chamber of the membrane dialysis reactor, d-1; q*FB,Glc – volume specific glucose uptake rate related to the fixed bed volume, mmol lFB
-1 h-1; q*FB,Gln – volume specific glutamine uptake rate related to the fixed bed volume, mmol lFB
-1 h-1. 相似文献