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.

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.

The production and effect of interferon on influenza virus growth in chick embryo lung epithelial and fibroblast cell cultures

1969 ◽  
Vol 15 (3) ◽  
pp. 273-277 ◽  
Author(s):  
Sunidhkumar S. Gandhi ◽  
Robert B. Stewart
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Cell Cultures ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Fibroblastic Cell ◽  
Virus Growth ◽  
Lung Epithelial ◽  
Fibroblastic Cells ◽  
Virus Strains

Cultures of fibroblastic cells prepared from chick embryo lung infected with low multiplicities of influenza type A virus strains were found to produce more interferon than did cultures of epithelial cells prepared from the same organ. Fibroblastic cell cultures were also found to be more sensitive to the action of interferon than were epithelial cells with respect to the levels of infectious virus produced and the duration of interferon action. Cultures of the two cell types treated with interferon did not differ with respect to the number of cells involved in virus synthesis.

The Immunohistochemical Characterisation of an SV40-immortalised Human Corneal Epithelial Cell Line

2003 ◽  
Vol 31 (4) ◽  
pp. 409-417 ◽  
Author(s):  
Anne Huhtala ◽  
Sami K. Nurmi ◽  
Hanna Tähti ◽  
Lotta Salminen ◽  
Päivi Alajuuma ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Cell Line ◽  
Cell Cultures ◽  
Corneal Epithelium ◽  
Culture Medium ◽  
Corneal Epithelial Cell ◽  
Cell Type ◽  
Corneal Epithelial ◽  
Single Cell Type

Alternatives to the Draize rabbit eye irritation test are currently being investigated. Because of morphological and biochemical differences between the rabbit and the human eye, continuous human cell lines have been proposed for use in ocular toxicology studies. Single cell-type monolayer cultures in culture medium have been used extensively in ocular toxicology. In the present study, an SV40-immortalised human corneal epithelial (HCE) cell line was characterised immunohistochemically, by using 13 different monoclonal antibodies to cytokeratins (CKs), ranging from CK3 to CK20. The results from the monolayer HCE cell cultures were compared with those from the corneal epithelium of human corneal cryostat sections. Previous studies have shown that the morphology of the HCE cell is similar to that of primary cultured human corneal epithelial cells, and that the cells express the cornea-specific CK3. In the study reported here, we show that the cell line also expresses CKs 7, 8, 18 and 19. These CKs are typically expressed by simple epithelial cells, and are not found in the human cornea in vivo. Therefore, the monolayer HCE cell line grown in culture medium does not express the CK pattern that is typical of human corneal epithelium. This should be taken into consideration when using HCE cell cultures in similar single cell-type experiments for ocular toxicology.

A method for measuring Cl efflux from dispersed cells of airway epithelium

1995 ◽  
Vol 78 (3) ◽  
pp. 1197-1202 ◽  
Author(s):  
T. Ohrui ◽  
B. Q. Shen ◽  
R. J. Mrsny ◽  
J. H. Widdicombe
Keyword(s):  
Cystic Fibrosis ◽  
Epithelial Cells ◽  
Room Temperature ◽  
Primary Cultures ◽  
Cell Types ◽  
Airway Epithelial Cells ◽  
Isolated Cells ◽  
T84 Cells ◽  
Tracheal Epithelial Cells ◽  
3T3 Fibroblasts

This paper describes a method for measuring the increase in halide permeability of isolated airway epithelial cells induced by adenosine 3′,5′-cyclic monophosphate (cAMP). Suspensions of isolated cells, known to contain the cystic fibrosis transmembrane conductance regulator (CFTR), were placed in the upper part of a Swinnex filter holder containing a filter with pores of 0.65 micron diameter. Medium was perfused over the cells at room temperature and collected at minute intervals following its passage through the filter. Experiments were performed on Calu-3 and T84 cells (human lung and colonic epithelial cell lines), primary cultures of dog and human tracheal epithelium, and Swiss 3T3 fibroblasts stably transfected with CFTR. In all cell types, addition of agents that elevate cAMP led to increases in the rates of loss of 36Cl and 125I. However, in human tracheal epithelial cells, warming the medium from room temperature to 37 degrees C was a more effective way of stimulating tracer efflux. Increases in efflux in response to either temperature or cAMP-elevating agents were inhibited by diphenylamine-2-carboxylate, a blocker of CFTR. Reproducible increases in tracer efflux were seen with as few as 10(6) cells. Cells that had been trypsinized off their culture dishes responded better than cells that had been scraped off, although treatment of scraped cells with trypsin enhanced their responsiveness to cAMP-elevating agents. Cystic fibrosis is characterized by the lack of a cAMP-activated Cl conductance in the apical membrane of airway epithlia.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Telomerase activity in primary cultures of normal adrenocortical cells

2001 ◽  
Vol 170 (3) ◽  
pp. 677-684 ◽  
Author(s):  
T Suwa ◽  
L Yang ◽  
PJ Hornsby
Keyword(s):  
Primary Culture ◽  
Adrenal Cortex ◽  
Cell Cultures ◽  
Primary Cultures ◽  
Telomerase Activity ◽  
Adrenocortical Cell ◽  
Isolated Cells ◽  
Adrenocortical Cells ◽  
Human Telomerase Reverse Transcriptase ◽  
In Cells

Telomerase activity was measured in isolated cells from bovine and human adrenal cortex, in cells in primary culture, in cells in later passages in culture, and in cells genetically modified by expression of hTERT (human telomerase reverse transcriptase). Telomerase activity in freshly isolated bovine adrenocortical cells and in human adrenal cells from donors of various ages (6-79 years) was very low or undetected. However, primary bovine adrenocortical cell cultures were strongly positive for telomerase activity, and primary human adrenocortical cell cultures were weakly positive. Both cell types proliferate in primary culture but proliferation of bovine cells is much more vigorous. When primary bovine cells were subcultured to make successively secondary and tertiary cultures, telomerase activity declined strongly, and was undetected by the third passage. There was only a slight decrease in growth rate over this period. Levels of the telomerase RNA component did not change with passage number when assessed by semi-quantitative competitive RT-PCR. When both bovine and human cells were infected with a retrovirus encoding hTERT, telomerase activity in the cells was very high. We conclude that in the adrenal cortex, as in some other tissues, TERT expression is regulated and upregulation of telomerase activity is associated with rapid proliferation in primary culture. Telomerase activity is not maintained, and introduction of TERT is required for stable telomerase activity and for immortalization.

Androgen metabolism and actions in rat ventral prostate epithelial and stromal cell cultures

1986 ◽  
Vol 64 (6) ◽  
pp. 583-593 ◽  
Author(s):  
J. Orlowski ◽  
A. F. Clark
Keyword(s):  
Epithelial Cells ◽  
Cell Cultures ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Cultured Cells ◽  
Cell Types ◽  
Ventral Prostate ◽  
Oxidoreductase Activity ◽  
Androgen Metabolism ◽  
Rat Ventral Prostate

The rat ventral prostate requires androgens for normal development, growth, and function. To investigate the relationship between androgen metabolism and its effects in the prostate and to examine differences between the epithelial and stromal cells, we have established a system of primary cell cultures of immature rat ventral prostate cells. Cultures of both cell types after reaching confluency (6–7 days) actively metabolized 3H-labelled testosterone (T), 5α-dihydrotestosterone (5α-DHT), 5α-androstane-3α,17β-diol, and 5α-androstane-3β,17β-diol. The epithelial cells actively reduced T to 5α-DHT and formed significant amounts of 5α-androstane-3,17-dione from T, 5α-DHT, and 5α-androstane-3α,17β-diol. All substrates were converted to significant amounts of C19O3metabolites. The stromal cells also metabolized all substrates, but very little 5α-androstane-3,17-dione was formed. The metabolism studies indicate that both cell types have Δ4-5α-reductase, 3α- and 3β-hydroxysteroid oxidoreductase and hydroxylase activities. The epithelial cells have significant 17β-hydroxysteroid oxidoreductase activity. The epithelial cells cultures grown in the presence of T have higher acid phosphatase (AP) contents (demonstrated histochemically and by biochemical assay). Tartrate inhibition studies indicate that the epithelial cells grown in the presence of T are making secretory AP. Stromal cell AP is not influenced by T. The results indicate that the cultured cells maintain differentiated prostatic functions: ability to metabolize androgens and, in the case of the epithelial cells, synthesize secretory AP.

scholarly journals Searching for new molecular markers for cells obtained from abdominal aortic aneurysm

2021 ◽  
Author(s):  
Marta Lesiak ◽  
Aleksandra Augusciak-Duma ◽  
Karolina L. Stepien ◽  
Agnieszka Fus-Kujawa ◽  
Malwina Botor ◽  
...  
Keyword(s):  
Gene Expression ◽  
Flow Cytometry ◽  
Cell Cultures ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Cell Types ◽  
Isolated Cells ◽  
Healthy Human ◽  
Relative Expression ◽  
And Control

AbstractThe aim of the study was to investigate specific potential markers for cells obtained from three layers of human AAA divided into three segments along the AAA based on morphological differences. The isolated cells were compared to control commercial cell types from healthy human abdominal aortas. For each type of aortic layer, three specimens from 6 patients were compared. Total RNA was isolated from 36 cell cultures for gene expression profiling and potential new cytometry markers were typed. Isolated cells were analyzed by flow cytometry by using fluorochrome-conjugated antibodies to markers: CNN1, MYH10, ENG, ICAM2, and TEK. The relative expression of 45 genes in primary cell cultures and control lines was analyzed. Statistically significant differences were found in the expression of most of the analyzed genes between individual layers and control lines. Based on relative expression, antibodies were selected for flow cytometry. Gene expression profiles allowed to select new potential cytometry markers: CNN1, MYH10, MYOCD, ENG, ICAM2, TEK. However, none of the tested markers seems to be optimal and characteristic for a specific layer of AAA.

scholarly journals Skin-resident immune cells actively coordinate their distribution with epidermal cells during homeostasis

2021 ◽  
Author(s):  
Sangbum Park ◽  
Catherine Matte-Martone ◽  
David G Gonzalez ◽  
Elizabeth A Lathrop ◽  
Dennis P May ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Density ◽  
Immune Cells ◽  
Immune Cell ◽  
Basal Layer ◽  
Cell Types ◽  
Epidermal Cells ◽  
Cell Junctions ◽  
Epithelial Stem Cells

Our organs consist of multiple cell types that ensure proper architecture and function. How different cell types coexist and interact to maintain their homeostasis in vivo remain elusive. The skin epidermis comprises mostly epithelial cells, but also harbors Langerhans cells (LCs) and Dendritic Epidermal T cells (DETCs). In response to injury or infection, LCs and DETCs become activated and play critical immunological roles. During homeostasis, they coexist with epithelial cells in the basal layer of the epidermis. Whether, and how, distributions of LCs and DETCs are regulated during homeostasis is unclear. Here, we addressed this question by tracking LCs, DETCs and epithelial basal cells over time within the skin of live adult mice. We show that LCs and DETCs maintain their overall position despite continuous turnover of neighboring basal epithelial stem cells. Moreover, LCs and DETCs rapidly and maximally explore basal epithelial cell junctions through their dendritic extensions. Altering the epithelial cell density triggers corresponding changes in the immune cell density, but not vice versa, suggesting that epithelial cells determine immune tissue composition in the epidermis. Moreover, LCs and DETCs are organized in a tiling pattern that is actively maintained. When LCs or DETCs are ectopically removed, neighboring epidermal LCs or DETCs, respectively, move into the emptied spaces and re-establish the tiling pattern. Finally, LCs require the GTPase Rac1 to maintain their positional stability, density and tiling pattern. Overall, we discovered that epidermal cells regulate the density of immune cells during homeostasis, and that immune cells actively maintain a non-random spatial distribution, reminiscent of neuronal self-avoidance. We propose that these cellular mechanisms provide the epidermis with an optimal response to environmental insults.

Kidney proximal tubule cells: Epithelial cells without EGTA-extractable annexins?

2000 ◽  
Vol 78 (4) ◽  
pp. 495-502 ◽  
Author(s):  
Sandra Tribolo ◽  
Suzanne Maroux ◽  
Dominique Massey-Harroche
Keyword(s):  
Epithelial Cells ◽  
Proximal Tubule ◽  
Primary Culture ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Isolated Cells ◽  
Vesicular Traffic ◽  
Kidney Proximal Tubule ◽  
Annexin I ◽  
Immunofluorescence Labelling

The expression and the subcellular localizations of annexins I, II, IV, VI, and XIII in renal epithelial cells were investigated, using immunological techniques with specific monoclonal antibodies. Upon performing Western blotting experiments, no annexins VI and XIII were detected in kidney, whereas annexins I, II, and IV were. Immunofluorescence labelling procedure performed on thin frozen renal sections showed the presence of these three annexins along the plasma membrane of the collecting duct cells with a restricted expression of annexin I at principal cells. Annexin I was also found present in some glomerular cells. None of these annexins, however, were detected in the proximal tubular cells upon performing immunofluorescence labelling and electrophoretic analysis on an EGTA (ethylenebis(oxyethylenenitrilo)tetraacetic acid)-extractable annexin fraction prepared from freshly isolated cells. This is the first time a mammalian epithelial cell has been found to express non-typical annexin (at least partly solubilized with EGTA). However, when these cells were grown in primary culture, they were found to express annexins I, II, IV, and V. As well as being located along the basolateral membrane, annexins I and II are also present on vesicles, which suggests that these annexins may be involved in vesicular traffic under cell culture conditions.Key words: annexin, kidney, proximal tubule, primary culture.

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.

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