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At least 865 records · Page 48Linked to original sources
PubMed1995

Two types of normal human breast epithelial cells derived from reduction mammoplasty: phenotypic characterization and response to SV40 transfection.

A culture method to grow two morphologically distinguishable normal human breast epithelial cell types derived from reduction mammoplasty has been developed. Type I cells were characterized by a more variable cell shape, smooth cell colony boundaries, the expression of epithelial membrane antigen (EMA) and keratin 18 and the non-expression of keratin 14 and alpha 6 integrin. In addition, the Type I cells were growth stimulated by fetal bovine serum (FBS) and were deficient in gap junctional intercellular communication (GJIC). In contrast, Type II cells were characterized by a uniform cell shape, expression of keratin 14 and alpha 6 integrin and the non-expression of EMA and keratin 18. In addition, Type II cells were growth inhibited by FBS and were proficient in GJIC. Type I cells can be induced by cholera toxin to change their morphology to a Type II cell morphology. Hence, Type I cells antigenically resemble luminal epithelial cells, while the Type II cells more closely resemble basal epithelial cells. Type I and Type II cells were transfected with SV40 DNA. Clones with extended lifespans were obtained from both Type I and Type II cells by SV40 transfection. Some (2/9) of the SV40-transfected Type I cell clones became immortal (> 100 cumulative population doubling level), whereas none (0/8) of the SV40-transfected Type II cell clones became immortal. The SV-40-transfected Type I and Type II cell-derived extended life clones and immortal cell lines phenotypically resembled their parental cells with respect to EMA, keratin 14 and keratin 18 expression and GJIC. Each (9/9) of the SV40 transfected Type I cell clones grew in soft agar; none (0/8) of the SV40-transfected Type II cell clones were capable of growing in soft agar. These results provide evidence that normal human breast epithelial cells, derived from reduction mammoplasty, can be separated into two morphologically and antigenically different cell types and that these two different cell types significantly differ in their response to an oncogenic (SV40) stimulus.

Breast↗
PubMedDate not supplied

Stimulation and inhibition of cell division in synchronized Escherichia coli.

Escherichia coli was synchronized by centrifugal elutriation. When grown in a Tris-based medium, addition of EDTA resulted in division about 20 min earlier (division of control at t = 75 min). EDTA addition caused a change in cell shape, with cells becoming narrower and longer, whereas the surface area to volume ratio increased. Irradiation with UV inhibited not only division and constriction, but also the increase in DAP incorporation found in dividing control cells. Possibly, division requires the construction of new polar caps, whereas premature division might involve remodeling of existing murein. In both cases, cell shape is presumed to be a relevant factor for division.

Cell Division↗
PubMed2003

Quantitative analysis of osteoblast behavior on microgrooved hydroxyapatite and titanium substrata.

The effects of implant surface topography and chemistry on osteoblast behavior have been a research focus because of their potential importance in orthopedic and dental applications. This work focused on the topographic effects of hydroxyapatite (HA) and titanium (Ti) surface that had identical micropatterns to determine whether there was synergistic interaction between surface chemistry and surface topography. Surface microgrooves with six different groove widths (4, 8, 16, 24, 30, and 38 microm) and three different groove depths (2, 4, and 10 microm) were made on single crystalline silicon wafers using microfabrication techniques. Ti and HA thin films were coated on the microgrooves by radio-frequency magnetron sputtering. After that, human osteoblast-like cells were seeded and cultured on the microgrooved surfaces for up to 7 days. The cells' behavior was examined using scanning electron microscopy after cells were fixed and dehydrated. Statistical analysis was based on quantitative data of orientation angle, evaluating the contact guidance, and form index, describing cell shape or cell morphology changes. The contact guidance and cell shape changes were observed on the HA and Ti microgrooves. No difference in orientation angle between HA and Ti microgrooves was found. This might suggest that surface chemistry was not a significant influence on cell guidance. However, the form index analysis indicated an interaction between topographic effects and surface chemistry. Thus, conclusions about surface topographic effects on cell behavior drawn from one type of material cannot simply be applied to another type of material.

Durapatite↗
PubMed2006

Pax6-induced alteration of cell fate: shape changes, expression of neuronal alpha tubulin, postmitotic phenotype, and cell migration.

The transcription factor Pax6 plays an important role in the development of the central nervous system. To understand its mechanism of action, we transduced HeLa cells with a Pax6-expressing lentiviral vector. Upon transduction, HeLa cells markedly changed shape and formed neuritelike extensions. Pax6-transduced HeLa cells expressed high levels of neuronal alpha3 tubulin, demonstrating a partial transdifferentiation towards a neuronal phenotype. Neurons are postmitotic cells. Pax6-transduced HeLa cells became postmitotic through mechanisms involving up-regulation of p53 and cyclin-dependent kinase inhibitor p21. One of the most striking effects of Pax6 was observed by time-lapse videomicroscopy: cells started to dissociate from cell clusters and displayed intense migratory activity. Migration was accompanied by dynamic and reversible shape changes. Our results identified three elements of Pax6 action: (i) expression of neuron-specific genes; (ii) establishment of a postmitotic phenotype; and (iii) involvement in the regulation of cell shape and cell migration.

Animals↗
PubMed1997

Epithelial folding in vitro: studies on the cellular mechanism underlying evagination of thyrocyte monolayers.

Epithelial monolayers in suspension culture fold in a way which closely resembles epithelial evagination. We have used freshly isolated segments of porcine thyroid follicles to study the mechanism underlying this evagination process. Epithelial folding was accompanied by dramatic changes in cell shape: the cells elongated and apical cell surfaces widened, whereas the basal cell portions were narrowed to about 20% of their original width. Apparently, enzymatic separation of thyroid epithelial cells from their underlying extracellular matrix resulted in an extension of the lateral cell-cell interactions on the expense of the basal cell surface area. Epithelial folding in vitro was Ca2+ dependent and reversibly blocked by cytochalasin D, by which the reorganization of the F-actin network was disturbed. This inhibitory effect was also observed by the action of cAMP analogues known to cause rounding of cells by their effect on cortical F-actin. Moreover, evagination in vitro was reversibly blocked at intracellular pH values of 5.8 and below. Under these conditions, protein phosphorylation was entirely inhibited. Inhibitors of protein kinases, specifically of myosin light chain kinase, were able to disrupt the evagination process, suggesting that protein phosphorylation, presumably of the myosin light chain, was essential for folding. We conclude that enzymatic separation of epithelial monolayers from their extracellular matrix initiated a cascade consisting of extended cell-cell interactions of the lateral plasma membranes and of reorganization of the apical actin-myosin network, finally resulting in profound changes in cell shape characteristic of epithelial evagination.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1989

Extracellular matrix organization modulates fibroblast growth and growth factor responsiveness.

To learn more about the relationship between extracellular matrix organization, cell shape, and cell growth control, we studied DNA synthesis by fibroblasts in collagen gels that were either attached to culture dishes or floating in culture medium during gel contraction. After 4 days of contraction, the collagen density (initially 1.5 mg/ml) reached 22 mg/ml in attached gels and 55 mg/ml in floating gels. After contraction, attached collagen gels were well organized; collagen fibrils were aligned in the plane of cell spreading; and fibroblasts had an elongated, bipolar morphology. Floating collagen gels, however, were unorganized; collagen fibrils were arranged randomly; and fibroblasts had a stellate morphology. DNA synthesis by fibroblasts in contracted collagen gels was suppressed if the gels were floating in medium but not if the gels were attached, and inhibition was independent of the extent of gel contraction. Therefore, growth of fibroblasts in contracted collagen gels could be regulated by differences in extracellular matrix organization and cell shape independently of extracellular matrix density. We also compared the responses of fibroblasts in contracted collagen gels and monolayer culture to peptide growth factors including fibroblast growth factor, platelet-derived growth factor, transforming growth factor-beta, and interleukin 1. Cells in floating collagen gels were generally unresponsive to any of the growth factors. Cells in attached collagen gels and monolayer culture were affected similarly by fibroblast growth factor but not by the others. Our results indicate that extracellular matrix organization influenced not only cell growth, but also fibroblast responsiveness to peptide growth factors.

Cell Adhesion↗
PubMed2005

Dynamics of temperature dependent modifications of blood in the near-infrared.

BACKGROUND AND OBJECTIVES: Laser irradiation induces blood coagulation by heating of blood components. It is a complex phenomenon which encompasses a variety of processes, such as cell shape modification, cell membrane rupture, protein denaturation, aggregation, and finally blood gelation. An in vitro study was performed to investigate heating temperatures leading to transformation of blood and to have a better understanding of the dynamics of temperature dependent modifications of blood in the near-infrared. STUDY DESIGN/MATERIALS AND METHODS: Slow heating of whole blood and hemolized blood was performed using a specific optical chamber. Eight hundred and five and 940 nm light transmission of blood was measured as a function of time during heating at various temperatures (60-75 degrees C). RESULTS: During heating of whole blood, three phases were clearly identified. For hemolized blood, only phase 3 was present. For whole blood, the duration of each phase was correlated to blood temperature. A temporary increase of transmission was observed during heating with a maximum at 65 degrees C. The analysis of the dynamics of temperature dependent modifications of blood are consistent with cell shape modification, denaturation, and aggregation of blood, resulting in the formation of an aqueous gel-matrix. CONCLUSIONS: "Slow" vessel heating which is now proposed as the optimal mechanism for permanent vessel closure should be reconsidered in terms of our results. In that particular case, the optical coefficients of blood could be different from those expected. In case of a large blood vessel, this could be an advantage since a "temporary" higher transmission of light could lead to a more homogeneous heating of the blood vessel. In case of a small blood vessel, this phenomenon could lead to a collateral damage of the skin.

Blood↗
PubMed2005

Prolonged unbalanced growth induces cellular senescence markers linked with mechano transduction in normal and tumor cells.

Cellular senescence is induced by diverse means and hence thought to be mediated by multiple pathways. We show that prolonged unbalanced growth due to retardation of DNA replication elicits a senescence-like phenomenon irrespective of the cell type. In fact, modest inhibition of DNA replication by various means led to cell swelling, cytoskeletal alterations, and irregularly enlarged, flat cell shape. Such cells upregulated senescence-associated genes, and eventually lost division potential. These phenotypes, which define cellular senescence, were virtually reversed by reducing protein synthesis or blocking ERK of the MAP kinase family. These results suggest that cellular senescence is a manifestation of prolonged unbalanced growth linked with mechano transduction and can be prevented by at least two different ways.

Actins↗
PubMed2004

Role of RhoA, mDia, and ROCK in cell shape-dependent control of the Skp2-p27kip1 pathway and the G1/S transition.

Cell shape-dependent control of cell-cycle progression underlies the spatial differentials of growth that drive tissue morphogenesis, yet little is known about how cell distortion impacts the biochemical signaling machinery that is responsible for growth control. Here we show that the Rho family GTPase, RhoA, conveys the "cell shape signal" to the cell-cycle machinery in human capillary endothelial cells. Cells accumulating p27(kip1) and arrested in mid G(1) phase when spreading were inhibited by restricted extracellular matrix adhesion, whereas constitutively active RhoA increased expression of the F-box protein Skp2 required for ubiquitination-dependent degradation of p27(kip1) and restored G(1) progression in these cells. Studies with dominant-negative and constitutively active forms of mDia1, a downstream effector of RhoA, and with a pharmacological inhibitor of ROCK, another RhoA target, revealed that RhoA promoted G(1) progression by altering the balance of activities between these two downstream effectors. These data indicate that signaling proteins such as mDia1 and ROCK, which are thought to be involved primarily in cytoskeletal remodeling, also mediate cell growth regulation by coupling cell shape to the cell-cycle machinery at the level of signal transduction.

Carrier Proteins↗
PubMed1991

[Identification of cytokines produced by cells cultured from human giant cell tumors of bone].

Morphological and biochemical studies were performed using tissue culture methods in 14 cases of giant cell tumor of bone. Primary cultures consisted of three types of cells, multinucleated giant cells, mononuclear round cells and spindle-shaped cells. The round cells were considered to be infiltrating macrophages, not neoplastic cells, according to the results obtained by morphological and immunohistochemical studies. The spindle-shaped cells were apparently neoplastic since they extensively proliferated and showed chromosomal abnormalities. Western blotting analysis of the supernatant fluid from the culture of the spindle-shaped cells showed the presence of several cytokines; M-CSF, IFN-gamma, TNF-alpha, which were known to show chemotactic, differentiation-inducing and activating effects on macrophages. When the conditioned mediums of cultured spindle-shaped cells were added to the culture of U-937 macrophage cell line, U-937 cells were induced to differentiate and became multinucleated giant cells. The results indicate that the spindle-shaped cells which could be passaged are neoplastic elements and that the cytokines produced by these cells have a significant role in clinicopathological status of the giant cell tumor of bone.

Adolescent↗
PubMed2006

Bacterial cell shape-dependent inflammatory response in mammary epithelial cells.

An in vitro co-culture model of SCp2 mammary epithelial cells and Escherichia coli strains was established in bacterial non-CO(2) incubators. Co-culturing SCp2 cells with either the rod-shaped W3110 or spherical-shaped GC7378Tn10 strains of Escherichia coli led to an increase in interleukin-6 (IL-6) levels by SCp2 cells after 9 h. At a ratio of 1:100 (epithelial:bacterial), the rod-shaped W3110 strain induced almost double the amount of IL-6 induced by the spherical-shaped GC7378Tn10 strain. The effect of Escherichia coli morphology (rod versus spherical) on IL-6 production by SCp2 cells was further investigated by shifting GC7378Tn10 morphology to rod through introducing the pbpA gene by transduction and transformation. In both approaches, the generated rod strains elicited higher IL-6 levels in SCp2 cells compared to the spherical ones at 1:50 and 1:100 ratios (epithelial:bacterial). Our findings demonstrate the significance of cell shape in bacterial-host interactions with potential implications in bacterial pathogenesis in general.

Animals↗
PubMed1981

Biochemistry of ATP-dependent red cell membrane shape change.

Red cell membranes prepared by hemolysis and washing in hypotonic Tris buffer crenate when suspended at 0 degrees C in isotonic medium. At 37 degrees C, in the presence of 1 mM MgATP, the crenated membranes are progressively converted to smooth-contoured discs and cup-forms. The phosphorylation of proteins and lipids during shape transformation in the presence of [gamma-32P]ATP has been studied. Spectrin phosphorylation and shape change could be dissociated in several ways, demonstrating that spectrin phosphorylation is neither necessary nor sufficient for the membrane smoothing reaction. Adenosine markedly inhibited phosphoinositide regeneration without altering shape change. Phosphatidic acid synthesis from endogenous diacylglycerol was not affected by adenosine and comparison of sheep, human and rabbit ghosts, which vary greatly in shape change capacity, demonstrated a direct correlation between phosphatidic acid synthesis and shape change rate. The results suggest that membrane curvature may be induced by diacyglycerol phosphorylation at the inner surface of the membrane bilayer, while the membrane skeleton limits the curvature and determines the shape ultimately assumed.

Adenosine↗
PubMed1997

Relationship between stiffness, internal cell pressure and shape of outer hair cells isolated from the guinea-pig hearing organ.

The mechanical properties of outer hair cells are of importance for normal hearing, and it has been shown that damage of the cells can lead to a reduction in the hearing sensitivity. In this study, we measured the stiffness of isolated outer hair cells in hyper- and hypotonic conditions, and examined the change in stiffness in relation to the corresponding changes in internal cell pressure and cell shape. The results showed that the axial stiffness of isolated outer hair cells (30-90 microns in length, 8-12 microns in diameter), ranging from 0.13-5.39 mN m-1, was inversely related to cell length. Exposure to hyper- and hypotonic external media with a small percentage change in osmolality caused a similar magnitude of change in cell length and cell diameter, but an average 60% change in cell stiffness. Therefore, a moderate osmotic change in the external medium can lead to a significant alteration in cell stiffness. The findings thus indicate an important contribution of internal cell pressure to cell stiffness.

Animals↗
PubMed1992

Intracellular Ca2+ concentration and pHi during metabolic inhibition.

To study the changes in intracellular Ca2+ concentration ([Ca2+]i) and pH (pHi) during metabolic inhibition, rat ventricular myocytes were dual loaded with the acetoxymethyl esters of fura-2 (fura-2/AM) and 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF/AM) and perfused with 2 mM sodium cyanide (NaCN). The percent of rod-shaped cells was 30% of the control after 30 min NaCN in the absence of glucose. [Ca2+]i increased from 82 +/- 8 to 151 +/- 25 (SE) nM (P less than 0.05) when cells were shortened, and to 421 +/- 106 nM (P less than 0.05) when cells were rounded. There was a positive relationship between pCai and pHi (r = 0.425, P less than 0.01). When 50 mM glucose was added during NaCN, there were no significant changes in [Ca2+]i and the percent of rod-shaped cells after 30 min. The pHi of rod-shaped cells, however, decreased to 6.95 +/- 0.03 (P less than 0.01). Although the acidic solution (70% O2-30% CO2) decreased pHi to 6.90 +/- 0.05 (P less than 0.01), there were no changes in the cell shape or [Ca2+]i. The addition of NaCN for 30 min decreased the rod-shaped cells to 18% of the control. Mild acidosis did not cause the changes in cell shape or [Ca2+]i. There was also no protection of the NaCN-induced changes in cell shape by intracellular acidosis. It is likely that the changes in cell shape during metabolic inhibition were related to the depletion of metabolic energy and the increase in [Ca2+]i.

Analysis of Variance↗
PubMed2003

[Distribution of microfilaments in detached corn root cap cells under fluorescent microscope].

The detached root cap cells from maize seedlings comprised of round-shaped cells, ellipse-shaped cells and longitude-shaped cells. By using fluorescein-iso-thiocyanate-phalloidin(FITC-Ph) as fluorescent probe and treatment with cytochalasin B (CB) or HMC toxin, a host-specific toxin from Bipolaris (Helminthosporium) maydis race C, the distribution and variation of microfilaments (MFs) in detached cells were investigated. The results were as follows (i) the round-shaped cells had intense fluorescence, but the network of MFs was not distinct. There was clear MFs network in the cytoplasm of both ellipse-shaped and longitude-shaped cells. The distribution of MFs in detached cells seemed to be relevant to their shape and vigor. (ii) the fluorescence of detached cells of Charrua cytoplasmic male sterility(cms-C) maize was decreased after treatment with HMC-toxin. The cause of this outcome was unclear. We only obtained the pictures of the distorted protoplast membrane and dead cells owing to treatment with HMC-toxin. The distribution of MFs of detached cells of Normal(N) cytoplasmic maize was not affected by HMC-toxin, and their protoplasts shank slightly. (iii) CB could change the distribution of MFs in detached cells of both cms-C maize and N maize to disordered arrangement.

Actin Cytoskeleton↗
PubMed2006

Pore structure of the packing of fine particles.

This paper presents a numerical study of the pore structure of fine particles. By means of granular dynamics simulation, packings of mono-sized particles ranging from 1 to 1000 microm are constructed. Our results show that packing density varies with particle size due to the effect of the cohesive van der Waals force. Pores and their connectivity are then analysed in terms of Delaunay tessellation. The geometries of the pores are represented by the size and shape of Delaunay cells and quantified as a function of packing density or particle size. It shows that the cell size decreases and the cell shape becomes more spherical with increasing packing density. A general correlation exists between the size and shape of cells: the larger the cell size relative to particle size, the more spherical the cell shape. This correlation, however, becomes weaker as packing density decreases. The connectivity between pores is represented by throat size and channel length. With decreasing packing density, the throat size increases and the channel length decreases. The pore scale information would be useful to understand and model the transport and mechanical properties of porous media.

Journal Article↗
PubMed1996

Shapes of bilayer vesicles with membrane embedded molecules.

The interdependence of the lateral distribution of molecules which are embedded in a membrane (such as integral membrane proteins) and the shape of a cell with no internal structure (such as phospholipid vesicles or mammalian erythrocytes) has been studied. The coupling of the lateral distribution of the molecules and the cell shape is introduced by considering that the energy of the membrane embedded molecule at a given site of the membrane depends on the curvature of the membrane at that site. Direct interactions between embedded molecules are not considered. A simple expression for the interaction of the membrane embedded molecule with the local membrane curvature is proposed. Starting from this interaction, the consistently related expressions for the free energy and for the distribution function of the embedded molecules are derived. The equilibrium cell shape and the corresponding lateral distribution of the membrane embedded molecules are determined by minimization of the membrane free energy which includes the free energy of the membrane embedded molecules and the membrane elastic energy. The resulting inhomogeneous distribution of the membrane embedded molecules affects the cell shape in a nontrivial manner. In particular, it is shown that the shape corresponding to the absolute energy minimum at given cell volume and membrane area may be elliptically non-axisymmetric, in contrast to the case of a laterally homogeneous membrane where it is axisymmetric.

Chemical Phenomena↗