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Results for “Cell Aggregation”

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At least 307 records · Page 17Linked to original sources

PubMed1998

Histodifferentiation of hair follicles in grafting of cell aggregates obtained by rotation culture of embryonic rat skin.

We have previously reported reconstruction of hair follicles from a single cell suspension of rat fetal upper lip by a two-step culture method consisting of rotation and flotation cultures. Rotation sorted out the cells and flotation facilitated histodifferentiation. In the present study, we added grafting procedures to the previous method to see whether cell aggregates obtained this way were graftable, and whether the grafting promoted histodifferentiation. The aggregates before and after flotation were grafted, and differentiation of hair follicles comparable to those in vivo was confirmed 10 days after grafting. There was no difference in the degree of differentiation between the two kinds of grafts. The grafting procedure therefore resulted in an appreciable increase in histodifferentiation even when aggregates obtained after flotation were grafted.

Animals↗

PubMed1995

Monocyte chemotactic protein-1 provokes mast cell aggregation and [3H]5HT release.

Monocyte chemotactic protein-1 (MCP-1) and MCP-3, the most active and representative compounds of the CC chemokine family, are proinflammatory cytokines that attract and activate specific types of leucocytes. We have used highly purified isolated rat peritoneal mast cells (RPMC) cultured for different lengths of time with and without MCP-1 (200, 100, 50 and 25 nM). Our data clearly show that MCP-1 (200 nM) causes a marked release of [3H]serotonin ([3H]5HT and histamine, which reach a peak at 40 min of incubation (56.6 +/- 5.3 and 34.7 +/- 6 above the control, respectively). In dose-response experiments, MCP-1 (200, 100, 50, 25, 12.5, 6.25 and 3.12 nM) provoked a dose-dependent release of [3H]5HT and histamine from RPMC, which was maximum at 200 nM. After preparation of the histidine decarboxylase (HDC) probe, a Northern blot analysis was determined for HDC mRNA. After 4 hr, steady-state levels of HDC mRNA were induced in a dose-dependent manner by MCP-1 (200-25 nM), compared to the controls. However, MCP-1 failed to prime RPMC in [3H]5HT and histamine release when C48/80 (0.05 micrograms/ml) or anti-IgE was used. In contrast, murine interleukin-3 (IL-3) in combination with MCP-1 (200 and 100 nM) provoked a greater release of histamine and [3H]5HT than the compounds alone. Moreover, RPMC treated with MCP-1 (200 nM) showed, under light microscopy (20x), greater clump formation, a phenomenon absent in the controls (untreated cells). The electron microscope studies revealed that treatment with MCP-1 (200 nM) promoted binding of RPMC and clearly demonstrated a communication between the cytoplasms of adjacent mast cells. Our report describes additional biological activities for MCP-1, suggesting for the first time that this human monocyte chemoattractant plays a fundamental role in histamine and serotonin release and cell aggregation in rat peritoneal mast cells.

Animals↗

PubMed1983

Red blood cell aggregation in nephrotic syndrome.

Red blood cell (RBC) aggregation induced by fibrinogen is a major determinant of the non-Newtonian flow behavior of human blood and has been suggested as a possible contributing factor for thrombogenesis. Given the elevated fibrinogen levels and the high incidence of thrombotic accidents in nephrotic syndrome (NS), a study was designed to assess RBC aggregation in this disease. Three different aggregation assays were used: (1) changes in light transmission through static cell suspensions, aggregation half time (AHT); (2) low gravity centrifugation of blood in nearly vertical tubes, zeta sedimentation ratio (ZSR); (3) direct observation of dilute RBC suspensions, microscopic aggregation index (MAI). The results indicate that RBC aggregation, as measured by AHT and ZSR, is increased significantly in NS patients versus healthy controls (P less than 0.001 for both methods); this increase was not a function of renal insufficiency. The AHT and ZSR data were well correlated with plasma fibrinogen levels (r = 0.75 and 0.84, respectively). No differences were observed with the MAI method, since with this technique the aggregation process reaches an asymptotic plateau for the range of fibrinogen found in NS. Implications of increased RBC aggregation and thus increased blood viscosity and flow disturbances on thrombogenesis are discussed.

Adolescent↗

PubMed1986

Phase resetting of spontaneously beating embryonic ventricular heart cell aggregates.

The influence of isolated 20-ms duration current pulses on the spontaneous rhythm of embryonic chick ventricular heart cell aggregates was studied. A pulse could either delay or advance the time of occurrence of the next action potential, depending on whether it fell early or late in the cycle. As the stimulus amplitude was increased, the transition from delay to advance occurred over a narrower range of coupling intervals. At low-stimulus amplitudes the transition from delay to advance occurred in a smooth continuous fashion; at medium-stimulus amplitudes the transition was discontinuous; at high-stimulus amplitudes graded action potentials were seen. It was impossible to annihilate spontaneous activity in aggregates with a single stimulus. The phase-resetting response to hyperpolarizing pulses was qualitatively the reverse of that produced by depolarizing pulses. A very high-amplitude depolarizing or hyperpolarizing pulse could produce rapid repetitive activity. Theoretical aspects of these phenomena are discussed.

Animals↗

PubMed1994

Changes of beta-amyloid precursor protein splice patterns in brain cell aggregate cultures.

The splice pattern of beta-amyloid precursor protein (beta-APP) has been studied in a variety of neuronal and glial cells and in brain cell aggregate cultures by the polymerase chain reaction (PCR). The brain-typical pattern, in which beta-APP695 is the dominant form, has been found only in aggregate cultures but not in any of the other cell types including neuronal cell lines. Selective elimination of glial cells from aggregates resulted in increased quantities of beta-APP695, whereas removal of neurons led to a reduction of beta-APP695 and to an elevation of beta-APP751 and beta-APP770. This shift of splice pattern was not observed in cocultures of the neuronal cell line PC 12 with primary astrocytes combined in a variety of cellular ratios. Blood serum, which is an essential component of these cultures, tested on aggregates, did not reduce the amount of beta-APP695 or have any marked effects on splice patterns generally. From these results it is concluded that investigations on brain-typical splicing of beta-APP require primary neurons. Neuronal cell lines may be no suitable model systems. Splicing events favoring production of beta-APP695 may mark an important, very early step of amyloid formation in the brain.

Amyloid beta-Protein Precursor↗

PubMed1984

Monitoring red blood cell aggregation with nuclear magnetic resonance.

We present a new application of nuclear magnetic resonance to monitor rouleaux formation in static blood suspensions of physiological hematocrit. The method measures the apparent mean residence time of water inside the rouleaux and yields the kinetics and the extent of red cell aggregation in units of cells per aggregate.

Adult↗

PubMed1985

Cellular uptake and subcellular distribution of mercury in chick embryonic retinal cell aggregates.

Cellular uptake and subcellular distribution of mercury (203HgCl2) were determined in the chick embryonic retinal cell aggregation system. The accumulation of mercury was dependent upon its concentration in the medium. The uptake was rapid; a maximum deposition of mercury at 5 microM occurred within the first 30 min followed by a decline. Accumulation of mercury at 1 microM was constant between 15 min and 24 hr. The subcellular distribution of mercury was observed in the following order: nuclei and cell debris greater than mitochondria-lysosomes greater than 105,000g supernatant greater than microsomes. The activity of acid phosphatase markedly decreased in the aggregates treated with mercury at 50 to 100 microM for 24 hr. Low concentrations of mercury at 0.5 to 5 microM showed an inhibition of this enzyme activity in a cell-free system. The results indicate a relationship between the amount of mercury in the cells and the toxicity it produced on the retinal cell aggregation system.

Acid Phosphatase↗

PubMed1996

Matrix-induced liver cell aggregates (MILCA) for bioartificial liver use.

Ex vivo reproduction of liver microstructure using isolated hepatocytes is critical for bioartificial liver use. We have developed a method of producing matrix-induced liver cell aggregates (MILCA) using a small number of collagen-coated beads as a nidus for formation of hepatocyte aggregates. Porcine hepatocytes were obtained by EDTA/collagenase digestion. Cell viability was assessed by trypan blue exclusion and LDH release. Cytochrome P-450 activity was determined at 4 and 24 hours by measuring the formation of 7-hydroxycoumarine (7-HC) from 7-ethoxycoumarine (7-EC). At 4 hours, the viability of MILCA was 92 +/- 2%, LDH release was 100 +/- 22 U/L and 7-HC formation was 140 +/- 34 nM/g cells. At 24 hours, MILCA viability remained greater than 90%, but 7-HC formation was lower than that of parallel control monolayer hepatocyte cultures (194 +/- 43 vs 481 +/- 78 nM/g cells; p < 0.002). On transmission electron microscopy, MILCA ultrastructure resembled that of a normal liver (maintenance of cell polarity, gap junctions, bile canaliculi, intact organellae, glycogen granules). MILCA were subsequently inoculated into hollow-fiber bioreactors which were perfused for 6 hours with plasma recovered from patients with fulminant hepatic failure (n = 6; 5 x 10(9) cells/cartridge, recirculation of 350 ml of plasma at 400 ml/min). In these studies, lidocaine (20 micrograms/ml) was cleared in less than 3 hours and 7-HC production at 6 hours was 71 +/- 8 nM/g cells. Other MILCA effects noted in this system included lowering of plasma lactate, bilirubin and ammonia and increase in the level of several non-essential amino acids.

Alginates↗

PubMed1991

Release of interleukin-6 from anterior pituitary cell aggregates: developmental pattern and modulation by glucocorticoids and forskolin.

The release of the immunologically active cytokine interleukin-6 (IL-6) by cultured anterior pituitary cell aggregates was found to increase with the age of the donor rats. The glucocorticoid hormone dexamethasone (DEX) dose-dependently inhibited the release of IL-6, the IC50 being 2.43 +/- 0.93 nM. The sex steroids dihydrotestosterone and estradiol were without detectable effect. The adenylate cyclase activator forskolin increased IL-6 release in aggregate cultures in the presence of DEX. It is concluded that IL-6 release by anterior pituitary cells is subject to developmental changes and under regulatory control of biologically active substances.

Adenylyl Cyclases↗

PubMed1985

From Beaumont to poison ivy: marine sponge cell aggregation and the secretory basis of inflammation.

We have studied Microciona prolifera cells as a model for inflammation and secretion. Dissociated in Ca-, Mg-free seawater with 2.5 mM EDTA, the cells aggregate when exposed to Ca (greater than 5 mM) and Ca ionophores. Extracellular Ca is not required over the course of aggregation; brief pulses of Ca suffice. Aggregation was induced by A23187 in excess EDTA after cells were prepared by pulse Ca. It appeared that Ca ionophore stimulated the secretion of Microciona aggregation factor (MAF) to a locus or in a form inaccessible to external EDTA. Pulse-induced aggregation depended on MAF because it was inhibited by MAF fragments, which are ligands for MAF-binding sites. Sponge cells were preloaded with three fluorescent dyes that monitor aspects of stimulus-secretion coupling: 1) 3,3'-dipropylthiadicarbocyanine iodide (dis-C3-(5)), a carbocyanine dye presumed to report changes in membrane potential; 2) 9-aminoacridine (9AA), which presumably reports secretion from acid vesicles; and 3) chlortetracycline (CTC), presumed to report mobilization of membrane-associated Ca. Exposure of cells either to constant Ca or to pulse Ca stimuli caused prompt decreases in the fluorescence of cells with diS-C3-(5) and increases in fluorescence of cells with 9AA. In contrast, although constant Ca provoked decreases in fluorescence of cells with CTC, a pulse Ca was without effect. Moreover, inhibitors of stimulus-response coupling (e.g., aspirin, sodium salicylate, 5 mM; diclofenac, 100 microM) inhibited sponge aggregation induced by either constant or pulse stimuli. In contrast, like the endogenous mediator of inflammation, leukotriene B4, trienoic alkyl catechols (urushiol) from poison ivy provoked aggregation. These studies suggest the utility of this marine model for analysis of stimulus-response coupling in cells of higher species that also respond to secretagogues in the absence of external Ca.

Aminacrine↗

PubMed1993

Beta-endorphin is a potent inhibitor of thymidine incorporation into DNA via mu- and kappa-opioid receptors in fetal rat brain cell aggregates in culture.

Thymidine incorporation into DNA was inhibited dose-dependently by beta-endorphin in rat fetal brain cell aggregate cultures. The inhibition was reversed partially by mu (cyclic D-Phe-Cys-Tyr-D-Trp-Orn-Thr- Pen-Thr amide) or kappa (norbinaltorphimine) antagonists. Complete blockade of the beta-endorphin inhibitory effect was achieved only on concomitant exposure to both antagonists. Eadie-Hofstee analysis revealed that beta-endorphin inhibited thymidine incorporation noncompetitively. In the presence of protease inhibitors, beta-endorphin decreased thymidine incorporation with an IC50 of 0.7 nM. Truncated and N-acetylated beta-endorphin derivatives, which bind with low affinity to opioid receptors, did not affect thymidine incorporation. These findings indicate that beta-endorphin at physiological concentrations can regulate thymidine incorporation in cultured brain cells.

Animals↗

PubMed2001

Parameters of red blood cell aggregation as correlates of the inflammatory state.

To identify clinically relevant parameters of red blood cell (RBC) aggregation, we examined correlations of aggregation parameters with C-reactive protein and fibrinogen in unstable angina (UA), acute myocardial infarction (AMI), and bacterial infection (BI). Aggregation parameters were derived from the distribution of RBC population into aggregate sizes (cells per aggregate) and changing of the distribution by flow-derived shear stress. Increased aggregation was observed in the following order: UA, AMI, and BI. The best correlation was obtained by integration of large aggregate fraction as a function of shear stress. To differentiate plasmatic from cellular factors in RBC aggregation, we determined the aggregation in the presence and absence of plasma and formulated a "plasma factor" (PF) ranging from 0 to 1. In AMI the enhanced aggregation was entirely due to PF (PF = 1), whereas in UA and BI it was due to both plasmatic and cellular factors (0 < or = PF < or = 1). It is proposed that clinically relevant parameters of RBC aggregation should express both RBC aggregate size distribution and aggregate resistance to disaggregation and distinguish between plasmatic and cellular factors.

Acute-Phase Reaction↗

PubMed1975

Microrheology and light transmission of blood. IV. The kinetics of artificial red cell aggregation induced by Dextran.

Employing both microscopic and photometric methods the rheology of pathological red cell aggregation was studied in model experiments. Suspensions of washed human red blood cells in dextran solutions containing rising concentrations of dextrans (M.W. 40000, 70000, 110000, 250000, 500000) were used. At low concentrations (less than 500 mg-%) of high molecular weight dextrans (greater than 70000) red cell suspensions formed aggregates similar to the ones found in normal human blood. At higher concentrations, the aggregates were similar to those observed in pathological human blood. The aggregates were studied under the condition of stasis, slow flow and at shear rate of their hydrodynamic dispersion. Besides, the flow behavior of the dispersed cells at high shear rates was studied. We found: 1. In all samples the rate of spontaneous aggregate re-formation in stasis (following hydrodynamic desaggregation) rose with rising dextran concentration up to 5.0 g-%. 2. The shear resistance of the aggregates, as measured by the shear stress necessary to keep them dispersed, rose up to concentrations of 2.5g-%, but fell at higher concentrations. 3. Only with dextran of a molecular weight above 110000 coarse agglomerates could be produced at high concentrations. Loose elastic meshes were rapidly produced at high concentrations of Dx 70. 4. When subjected to steady state low shear (m sec-1) only the agglomerates, but not the meshes rapidly grew in size. Most of the aggregation kinetics recorded by photometry and microscopy evaded detection by viscometry.

Blood Viscosity↗

PubMed2004

Analysis of red blood cell aggregation in cardio-pulmonary bypass (CPB) surgery.

Not much is known about red cell aggregation during cardio-pulmonary bypass surgery (CPB). Blood samples from 19 patients undergoing CPB were anticoagulated with EDTA. Hematocrit was adjusted to 40%. A red blood cell aggregometer (LORCA) measured changes in light reflection from each blood sample after cessation of the rotation, and calculated an aggregation index (AI). Reflection measurements were stored. Because LORCA software failed for 87 of 171 samples, we developed new software, and applied it to the stored reflection measurements. This software failed only in 7 out of 171 cases and showed that all LORCA failures occurred for AI < 40%. The new calculations revealed that the aggregation index significantly decreased from 46.6 +/- 10.1 (mean +/- standard deviation) baseline to 22.8 +/- 8.3 at the end of CPB and recovered to 37.1 +/- 13.5 at day 1. It is concluded that the new software can be used to study decreased red cell aggregation during CPB.

Coronary Artery Bypass↗

PubMed2006

Increased red blood cell aggregation in patients with idiopathic intracranial hypertension. A hitherto unexplored pathophysiological pathway.

It was the objective of this study to explore the possibility that the aggregation of red blood cells is enhanced in individuals with increased intracranial hypertension (IIH). This is a prospective cross sectional examination in a cohort of patients with IIH and matched controls. The aggregation of red blood cells in the peripheral venous blood was determined by using a slide test and image analysis. We have presently included a group of 33 women with IIH and the same number of women matched for age, body mass index, vascular risk factors and medications. A significant (p = 0.038) increment in fibrinogen concentrations was noted in the patients (341 +/- 60.8 mg/dl) as opposed to the controls 307.9 +/- 64.8). The same stands for the aggregation of red blood cells (aggregation parameter of 8.7 +/- 4.9 in patients vs.5.9 +/- 3.2 in the controls, p = 0.001). We noted an increment in the aggregation of red blood cells in the peripheral blood of 33 women with IIH as opposed to matched controls. Being associated with capillary slow flow, these findings might be relevant to the ethiopathogenesis of this disease.

Adolescent↗

PubMed1977

The influence of light and different ATP concentrations on cell aggregation in cyclic AMP sensitive and insensitive species of the cellular slime molds.

The influence of light and different concentrations of ATP on cell aggregation in cyclic AMP sensitive (Dictyostelium mucoroides, D. purpureum) and cyclic AMP insensitive species (Polysphondylium violaceum, P. pallidum, D. lacteum) of the cellular slime molds was observed in small and in large amoebal populations. Both light and ATP (optimal concentration: 10(-5) M) accelerated cell aggregation and increased the number of aggregating centers in large populations. For cyclic AMP sensitive species the effect of ATP in large populations was more pronounced than for the species that do not react to cyclic AMP. A possible explanation for the similar effect of light and ATP has been discussed.

Adenosine Triphosphate↗

PubMed1988

Nevus cell aggregates in the placenta. A histochemical and electron microscopic study.

This is a report of the histochemical and ultrastructural analysis of nevus cell aggregates in the placenta from a neonate with a giant pigmented nevus. Although the distinction between aberrant migration of neural crest elements and blood-borne metastasis from the giant pigmented nevus is not readily resolved, the authors favor the former as an explanation of this unusual placental lesion.

Biopsy↗

PubMed1997

Effect of POMC(1-76), its C-terminal fragment gamma3-MSH and anti-POMC(1-76) antibodies on DNA replication in lactotrophs in aggregate cell cultures of immature rat pituitary.

Treatment of aggregate cell cultures of 14-day-old rat pituitary for 40 h with purified human (h) POMC(1-76) dose-dependently augmented the number of DNA replicating lactotrophs as estimated by autoradiography of [3H]-thymidine (3H-T) incorporation in cells immunostained for prolactin (PRL). No such effect was seen on the total number of 3H-T labelled cells (the majority of which did not contain any pituitary hormone in a detectable amount) or on the total number of lactotrophs. The effect of hPOMC(1-76) on 3H-T incorporation in lactotrophs was blocked by concomitant treatment with anti-hPOMC(1-76) monoclonal and polyclonal antibodies cross-reactive with rat POMC(1-74). The latter anti-hPOMC(1-76) antibodies also decreased the number of 3H-T incorporating lactotrophs in the absence of hPOMC(1-76). Gamma3-MSH, which is the C-terminal domain of hPOMC(1-76), mimicked the effect of hPOMC(1-76) on 3H-T incorporation in lactotrophs but its potency was lower than that of hPOMC(1-76). Other melanocortin (MC) peptides such as alpha- and beta-MSH were also effective but were less potent than gamma3-MSH. The difference in potency was not due to partial degradation of the peptides. hPOMC(1-76) did not affect 3H-T incorporation in other pituitary cell types. In contrast gamma3-MSH also augmented the number of 3H-T labelled somatotrophs and thyrotrophs. In the embryonic kidney 293 cell line stably transfected with the MC-3 receptor, gamma3-MSH (10 nM) augmented cAMP formation up to 30 times. In contrast, hPOMC(1-76) (100 nM) was inactive in this test system, indicating this peptide is not an agonist at the MC-3 receptor. The present investigation further supports the role of rat POMC(1-74) as a paracrine growth factor in the development of lactotrophs. The active core of POMC(1-76) does not seem to be restricted to its C-terminal domain gamma3-MSH as the latter peptide displays a growth promoting effect that is different from that of POMC(1-76): it is less potent, it is not specific for lactotrophs and whereas the effect of gamma3-MSH may be mediated by the MC-3 receptor that of POMC(1-76) is not.

Animals↗