Rapid cellular translocation is related to close contacts formed between various cultured cells and their substrata

1982 ◽  
Vol 54 (1) ◽  
pp. 23-34
Author(s):  
J. Kolega ◽  
M.S. Shure ◽  
W.T. Chen ◽  
N.D. Young
Keyword(s):  
Cultured Cells ◽  
Mdck Cells ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Cellular Translocation ◽  
Focal Contacts ◽  
Contact Patterns ◽  
Chick Heart ◽  
Close Contacts

Interference-reflection microscopy combined with time-lapse cinemicrography was used to examine the relationship between cell-to-substratum contact patterns and the speeds of translocation for a variety of cell types. Rapid translocation of amphibian leukocytes (average speed = 9.0 micron/min), amphibian epidermal cells (7 micron/min) and teleost epidermal cells (7 micron/min) was found to correlate with patterns of broad grey close contacts. Similar contact patterns were found under freshly seeded (2 h) chick heart fibroblasts (moving 1–3 micron/min), the rapidly advancing (1-5 micron/min) margin of spreading human WI-38 fibroblasts, and isolated MDCK canine epithelial cells (0.5-1.0 micron/min). Conversely, numerous dark streaks of focal contact were found associated with the slow rate of translocation displayed by older cultures (72 h) of chick fibroblasts (less than 0.1 micron/min), well-spread WI-38 cells (less than or equal to 0.3 micron/min) and confluent MDCK cells (less than 0.01 micron/min). It is concluded that close contacts, but not focal contacts, are associated with rapid cellular translocation, and that the build-up of focal contacts is associated with reduced cellular translocation and maintenance of the spread cell shape.

Contact behaviour during the reassociation of dissociated epithelial cells in primary culture

1987 ◽  
Vol 88 (4) ◽  
pp. 521-526
Author(s):  
R.M. Brown ◽  
C.A. Middleton
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Primary Culture ◽  
Cell Cultures ◽  
Corneal Epithelium ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Isolated Cells ◽  
Contact Behaviour

The behaviour in culture of dissociated epithelial cells from chick embryo pigmented retina epithelium (PRE), corneal epithelium (CE) and epidermis has been studied using time-lapse cinematography. The analysis concentrated on the contact behaviour of 60 previously isolated cells of each type during a 24 h period starting 3.5 h after the cells were plated out. During the period analysed the number of isolated cells in cultures of all three types gradually decreased as they became incorporated into islands and sheets of cells. However, there were significant differences in behaviour between the cell types during the establishment of these sheets and islands. In PRE cell cultures, islands of cells developed because, throughout the period of analysis, collisions involving previously isolated cells almost invariably resulted in the development of a stable contact. Once having established contact with another cell these cells rarely broke away again to become reisolated. In contrast the contacts formed between colliding CE and epidermal cells were, at least initially, much less stable and cells of both these types were frequently seen to break away and become reisolated after colliding with other cells. Sheets and islands of cells eventually developed in these cultures because the frequency with which isolated cells become reisolated decreased with increasing time in culture. The possible reasons underlying the different behaviour of PRE cells, when compared with that of CE and epidermal cells, are discussed. It is suggested that the decreasing tendency of isolated CE and epidermal cells to become reisolated may be related to the formation of desmosomes.

scholarly journals Invasive behaviour of mouse primordial germ cells in vitro

1986 ◽  
Vol 86 (1) ◽  
pp. 133-144
Author(s):  
D. Stott ◽  
C.C. Wylie
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Time Lapse ◽  
Cell Types ◽  
Fluorescent Labelling ◽  
Mouse Embryo Fibroblast ◽  
Substrate Interaction ◽  
Close Contacts

We have isolated migrating primordial germ cells (PGCs) from 10.5-day mouse embryos and studied their behaviour when cultured on a mouse embryo fibroblast (STO) cell line. Living and fixed PGCs were identified by fluorescent labelling with a monoclonal antibody specific for PGCs in the culture system used. The behaviour of the cells was studied using interference reflexion microscopy (IRM) and time-lapse video cinematography. The IRM pattern displayed by PGCs is typical of highly motile cell types, the cells lack focal contacts and possess large areas of close contacts indicative of weak membrane to substrate interaction. The PGCs exhibit relatively high rates of translocation and lack contact inhibition. They were observed to underlap STO cells in subconfluent monolayers and to penetrate between the cells of confluent monolayers, becoming located between the monolayer and its substrate. These observations support the hypothesis that migrating mouse PGCs are inherently motile and are able transiently to disrupt the adhesion of surrounding cells. These results suggest that PGCs actively migrate to the developing gonad in vivo.

Locomotory behaviour of epitheliocytes and fibroblasts on metallic grids

1999 ◽  
Vol 112 (8) ◽  
pp. 1273-1282
Author(s):  
Y.A. Rovensky ◽  
L.V. Domnina ◽  
O.Y. Ivanova ◽  
J.M. Vasiliev
Keyword(s):  
Mdck Cells ◽  
Time Lapse ◽  
Cell Types ◽  
Human Foreskin Fibroblasts ◽  
Primary Mouse Hepatocytes ◽  
Primary Mouse ◽  
Rat Liver Cells ◽  
Mouse Hepatocytes ◽  
Metallic Grids ◽  
Locomotory Behaviour

Behaviour of epitheliocytes and fibroblasts on special discontinuous substrata (metallic grids with square openings of 45x45 microm2) was examined in order to compare the ability of these cells to spread in two mutually perpendicular directions and to stretch over the void spaces. Two cell types with typical fibroblastic morphology, the AGO 1523 line of human foreskin fibroblasts and secondary cultures of mouse embryo fibroblasts, and three cell types with typical epithelial morphology, primary mouse hepatocytes, the IAR-2 line of rat liver cells and the MDCK line of canine kidney epithelial cells (clone 20) were used. We also examined the epitheliocytes (MDCK cells, clone 20) transformed to fibroblast-like morphology by treatment with hepatocyte growth factor/scatter factor (HGF/SF). Time-lapse video microscopy, scanning electron microscopy and immunofluorescence microscopy were used to examine cell reorganizations at various stages of spreading. It was found that early stages of spreading of fibroblasts and epitheliocytes were similar: the cell spread along two bars, perpendicular to each other (bar and crossbar), with the formation of a small triangular lamellar cytoplasm stretched over the opening. Later central parts of the bodies of the fibroblasts retracted from the bars so that the cells remained attached only by their polar lamellae. Successive expansions and partial retractions of these lamellae led to elongation of the cell body crossing several openings of the grid. Epitheliocytes, in contrast to fibroblasts, at the late stages of spreading did not retract their bodies and did not contract polar lamellae. As a result, their central lamellae stretched progressively over the openings. As a result of the treatment of MDCK epitheliocytes with HGF/SF the behaviour of the cells on the grids became similar to that of fibroblasts. It is suggested that these distinct spreading patterns of epitheliocytes and fibroblasts are due to the type-specific differences in the actin-myosin cortex. Experiments with microtubule-specific drugs, colcemid and taxol, indicate that the organization of this cortex is under microtubular control.

Morphology and locomotion of individual epithelial cells in culture

1985 ◽  
Vol 78 (1) ◽  
pp. 105-115 ◽  
Author(s):  
R.M. Brown ◽  
C.A. Middleton
Keyword(s):  
Epithelial Cells ◽  
Primary Cultures ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Cell Type ◽  
Isolated Cells ◽  
Cell Behaviour ◽  
Significant Movement ◽  
Different Cell Types

The behaviour in culture of epithelial cells derived from chick embryo pigmented retina epithelium (PRE), corneal epithelium (CE) and epidermis has been studied using time-lapse cinemicrography. The analysis concentrated on the morphology and movement of individual isolated cells, lacking contacts with other cells, during a 24h period starting 1–3 h after the cells were plated out in primary cultures. Isolated cells from all three sources could change morphology and reversibly exhibited either a poorly spread or a well-spread morphology. While poorly spread, the different cell types all appeared similar and all blebbed vigorously. In contrast, while well spread, the cells did not bleb significantly but there were other differences between them. Well-spread CE cells were always polarized by the presence of a dominant leading lamella but well-spread PRE cells were always unpolarized. Well-spread epidermal cells exhibited both a polarized and an unpolarized morphology. The tendency of individual isolated cells to change morphology varied with cell type. PRE cells were the most stable. Nearly 80% of them retained the same morphology throughout the period of analysis and only 1% of them showed three or more changes in morphology during this period. In contrast, 22% of CE cells and 37% of epidermal cells showed three or more changes in morphology during the period of observation. Isolated cells of all three types spent a greater proportion of the time exhibiting a poorly spread morphology than they spent exhibiting any alternative well-spread morphology. The analysis revealed a relationship between the morphology of isolated cells and the speed of their locomotion. Only cells with a well-spread polarized morphology showed significant movement. CE and epidermal cells with this morphology moved three to four times faster than their counter-parts with a poorly spread morphology or, in the case of epidermal cells, with a well-spread but unpolarized morphology. Actively moving PRE cells were not seen and this correlates with the absence of cells with a well-spread polarized morphology from cultures of this type. These findings are discussed in the light of similar investigations of cell behaviour in other epithelial cell types and fibroblasts.

Contacts of chick fibroblasts on glass: results and limitations of quantitative interferometry

1988 ◽  
Vol 90 (2) ◽  
pp. 215-224
Author(s):  
J. Bailey ◽  
D. Gingell
Keyword(s):  
Refractive Index ◽  
Cell Surface ◽  
Limb Bud ◽  
Membrane Bilayer ◽  
Focal Contacts ◽  
Wide Range ◽  
Chick Heart ◽  
Close Contacts ◽  
Migrating Cells

We have examined the contacts made by explanted chick heart and limb bud fibroblasts after 24–48 h on glass, using quantitative interference reflection microscopy (IRM). Contacts beneath very thin cytoplasmic lamellae were avoided because the images of such contacts depend on the thickness of the lamellae. Plaque-like focal contacts, distinguished on the basis of shape and low irradiance (darkness), are intimate adhesions to the substratum. These images can be interpreted if it is assumed that microfilaments associated with the lower membrane increase the local cytoplasmic refractive index. The range of irradiances measured for focal contacts was found to be rather wide, and our modelling shows that the most likely explanation for this is that the images receive variable contributions from the adjacent cytoskeleton. For this reason it is particularly difficult to assign a characteristic thickness for these contacts from IRM data. Close contacts, seen principally as ‘grey’ regions under migrating cells at the edges of the explants, also show a wide range of irradiances. Unlike focal contacts, it is not necessary to postulate any involvement of the cytoskeleton in their images and they can be modelled as regions where an aqueous glycocalyx zone about 20–30 nm thick separates the membrane bilayer from the glass. Paler grey regions that also look like close contacts are apparently formed where the cell surface has lifted several tens of nanometres from the glass.

A Deficiency Screen for Zygotic Loci Required for Establishment and Patterning of the Epidermis in Caenorhabditis elegans

Genetics ◽  
1997 ◽  
Vol 146 (1) ◽  
pp. 185-206 ◽  
Author(s):  
Rebecca M Terns ◽  
Peggy Kroll-Conner ◽  
Jiangwen Zhu ◽  
Sooyoun Chung ◽  
Joel H Rothman
Keyword(s):  
Caenorhabditis Elegans ◽  
Cell Types ◽  
Epidermal Cells ◽  
Epidermal Differentiation ◽  
Apparent Effect ◽  
Internal Organs ◽  
C Elegans ◽  
Seam Cell ◽  
Genomic Regions ◽  
Caenorhabditis Elegans Genome

To identify genomic regions required for establishment and patterning of the epidermis, we screened 58 deficiencies that collectively delete at least ∼67% of the Caenorhabditis elegans genome. The epidermal pattern of deficiency homozygous embryos was analyzed by examining expression of a marker specific for one of the three major epidermal cell types, the seam cells. The organization of the epidermis and internal organs was also analyzed using a monoclonal antibody specific for epithelial adherens junctions. While seven deficiencies had no apparent effect on seam cell production, 21 were found to result in subnormal, and five in excess numbers of these cells. An additional 23 deficiencies blocked expression of the seam cell marker, in some cases without preventing cell proliferation. Two deficiencies result in multinucleate seam cells. Deficiencies were also identified that result in subnormal numbers of epidermal cells, hyperfusion of epidermal cells into a large syncytium, or aberrant epidermal differentiation. Finally, analysis of internal epithelia revealed deficiencies that cause defects in formation of internal organs, including circularization of the intestine and bifurcation of the pharynx lumen. This study reveals that many regions of the C. elegans genome are required zygotically for patterning of the epidermis and other epithelia.

Applying Probe Electrospray Ionization Mass Spectrometry to Cytological Diagnosis: A Preliminary Study by Using Cultured Lung Cancer Cells

2021 ◽  
pp. 1-10
Author(s):  
Yuki Morimoto ◽  
Takeshi Oya ◽  
Mayuko Ichimura-Shimizu ◽  
Minoru Matsumoto ◽  
Hirohisa Ogawa ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electrospray Ionization ◽  
Squamous Cell ◽  
Cultured Cells ◽  
Sample Pretreatment ◽  
Cell Types ◽  
Diagnostic Tools ◽  
Ionization Mass ◽  
Diagnostic Modality ◽  
Probe Electrospray Ionization

<b><i>Objectives:</i></b> Cytology and histology are 2 indispensable diagnostic tools for cancer diagnosis, which are rapidly increasing in importance with aging populations. We applied mass spectrometry (MS) as a rapid approach for swiftly acquiring nonmorphological information of interested cells. Conventional MS, which primarily rely on promoting ionization by pre-applying a matrix to cells, has the drawback of time-consuming both on data acquisition and analysis. As an emerging method, probe electrospray ionization-MS (PESI-MS) with a dedicated probe is capable to pierce sample and measure specimen in small amounts, either liquid or solid, without the requirement for sample pretreatment. Furthermore, PESI-MS is timesaving compared to the conventional MS. Herein, we investigated the capability of PESI-MS to characterize the cell types derived from the respiratory tract of human tissues. <b><i>Study Design:</i></b> PESI-MS analyses with DPiMS-2020 were performed on various type of cultured cells including 5 lung squamous cell carcinomas, 5 lung adenocarcinomas, 5 small-cell carcinomas, 4 malignant mesotheliomas, and 2 normal controls. <b><i>Results:</i></b> Several characteristic peaks were detected at around m/z 200 and 800 that were common in all samples. As expected, partial least squares-discriminant analysis of PESI-MS data distinguished the cancer cell types from normal control cells. Moreover, distinct clusters divided squamous cell carcinoma from adenocarcinoma. <b><i>Conclusion:</i></b> PESI-MS presented a promising potential as a novel diagnostic modality for swiftly acquiring specific cytological information.

Quantitative Studies of Fibronectin Adsorption on Submicron Scaffolds

2012 ◽  
Author(s):  
Shawn Regis ◽  
Manisha Jassal ◽  
Sina Youssefian ◽  
Nima Rahbar ◽  
Sankha Bhowmick
Keyword(s):  
Surface Functionalization ◽  
Cultured Cells ◽  
Cell Attachment ◽  
Md Simulations ◽  
Cell Types ◽  
Biological Processes ◽  
Integrin Receptors ◽  
Tissue Engineering Scaffolds ◽  
Quantitative Studies

Fibronectin plays a crucial role in adhesion of several cell types, mainly due to the fact that it is recognized by at least ten different integrin receptors. Since most cell types can bind to fibronectin, it becomes involved in many various biological processes. The interaction of cells with ECM proteins such as fibronectin provides the signals affecting morphology, motility, gene expression, and survival of cells [1]. Fibronectin exists in both soluble and insoluble forms; soluble fibronectin is secreted by cells and exits in cell media or body fluids, whereas insoluble fibronectin exists in tissues or the extracellular matrix of cultured cells [2]. The ability to control adsorption of fibronectin on tissue engineering scaffolds would therefore play a huge role in controlling cell attachment and survival in vivo. This can be achieved through surface functionalization of the scaffolds. The goal of these studies is to use molecular dynamics (MD) simulations to mechanistically understand how fibronectin adsorption is enhanced by surface functionalization of submicron scaffolds.

scholarly journals Impact of Heterogeneity within Cultured Cells on Bacterial Invasion: Analysis of Pseudomonas aeruginosa andSalmonella enterica Serovar Typhi Entry into MDCK cells by Using a Green Fluorescent Protein-Labelled Cystic Fibrosis Transmembrane Conductance Regulator Receptor

2000 ◽  
Vol 68 (2) ◽  
pp. 861-870 ◽  
Author(s):  
A. Alev Gerçeker ◽  
Tanweer Zaidi ◽  
Peter Marks ◽  
David E. Golan ◽  
Gerald B. Pier
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Fluorescent Protein ◽  
Cultured Cells ◽  
Mdck Cells ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Bacterial Uptake ◽  
Cftr Protein ◽  
Green Fluorescent

ABSTRACT The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride ion channel that also serves as a receptor for entry ofPseudomonas aeruginosa and Salmonella entericaserovar Typhi into epithelial cells. To evaluate heterogeneity in CFTR protein expression in cultured cells and the effect of heterogeneity on internalization of different P. aeruginosa and serovar Typhi strains, we used two-color flow cytometry and confocal laser microscopy to study bacterial uptake by Madin-Darby canine kidney (MDCK) type I epithelial cells stably expressing a green fluorescent protein (GFP)-CFTR fusion construct (MDCK–GFP-CFTR cells). We found a strong correlation between cell size and GFP-CFTR protein expression, with 60 to 70% of cells expressing low levels of GFP-CFTR protein, 20 to 30% expressing intermediate levels, and <10% expressing high levels. The cells were sorted into low-, intermediate-, or high-level producers of CFTR protein; in vitro growth of each sorted population yielded the same distribution of CFTR protein expression as that in the original population. Cells expressing either low or high levels of CFTR protein internalized bacteria poorly; maximal bacterial uptake occurred in the cells expressing intermediate levels of CFTR protein. Treatment of MDCK cells with sodium butyrate markedly enhanced the production of CFTR protein without increasing cell size; butyrate treatment also increased the proportion of cells with internalized bacteria. However, there were fewer bacteria per butyrate-treated cell and, for P. aeruginosa, there was an overall decrease in the total level of bacterial uptake. The most highly ingested bacterial strains were internalized by fewer total MDCK–GFP-CFTR cells, indicating preferential bacterial uptake by a minority of epithelial cells within a given culture. Confocal fluorescence microscopy showed that P. aeruginosa and serovar Typhi induced cytoplasmic accumulation of CFTR protein close to the plasma membrane where the bacteria were adherent. These results show that within a population of MDCK–GFP-CFTR cells, there are cells with markedly different abilities to ingest bacteria via CFTR, the majority of the P. aeruginosa and serovar Typhi cells are ingested by the one-fourth to one-third of the cells that exhibit an intermediate size and level of CFTR protein expression, and overexpression of the CFTR receptor does not increase total bacterial uptake but rather allows more epithelial cells to ingest fewer total bacteria.

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