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.

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 EEA1, a Tethering Protein of the Early Sorting Endosome, Shows a Polarized Distribution in Hippocampal Neurons, Epithelial Cells, and Fibroblasts

2000 ◽  
Vol 11 (8) ◽  
pp. 2657-2671 ◽  
Author(s):  
Jean M. Wilson ◽  
Meltsje de Hoop ◽  
Natasha Zorzi ◽  
Ban-Hock Toh ◽  
Carlos G. Dotti ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mammalian Cells ◽  
Hippocampal Neurons ◽  
Cell Types ◽  
Early Endosomes ◽  
Filamentous Material ◽  
Endocytic Vesicles ◽  
Fibroblastic Cells ◽  
Darby Canine Kidney ◽  
Endocytic Pathways

EEA1 is an early endosomal Rab5 effector protein that has been implicated in the docking of incoming endocytic vesicles before fusion with early endosomes. Because of the presence of complex endosomal pathways in polarized and nonpolarized cells, we have examined the distribution of EEA1 in diverse cell types. Ultrastructural analysis demonstrates that EEA1 is present on a subdomain of the early sorting endosome but not on clathrin-coated vesicles, consistent with a role in providing directionality to early endosomal fusion. Furthermore, EEA1 is associated with filamentous material that extends from the cytoplasmic surface of the endosomal domain, which is also consistent with a tethering/docking role for EEA1. In polarized cells (Madin-Darby canine kidney cells and hippocampal neurons), EEA1 is present on a subset of “basolateral-type” endosomal compartments, suggesting that EEA1 regulates specific endocytic pathways. In both epithelial cells and fibroblastic cells, EEA1 and a transfected apical endosomal marker, endotubin, label distinct endosomal populations. Hence, there are at least two distinct sets of early endosomes in polarized and nonpolarized mammalian cells. EEA1 could provide specificity and directionality to fusion events occurring in a subset of these endosomes in polarized and nonpolarized cells.

scholarly journals 3D printed lung on a chip device with a stretchable nanofibrous membrane for modeling ventilator induced lung injury

2021 ◽  
Author(s):  
Sinem Tas ◽  
Emil Rehnberg ◽  
Deniz A. Bölükbaş ◽  
Jason P. Beech ◽  
Liora Nasi Kazado ◽  
...  
Keyword(s):  
Cell Death ◽  
Epithelial Cells ◽  
Lung Injury ◽  
Mechanical Stretch ◽  
Cell Types ◽  
Lung Epithelial Cells ◽  
Nanofibrous Membrane ◽  
Ventilator Induced Lung Injury ◽  
Lung Epithelial ◽  
3D Printed

Mechanical ventilation is often required in patients with pulmonary disease to maintain adequate gas exchange. Despite improved knowledge regarding the risks of over ventilating the lung, ventilator induced lung injury (VILI) remains a major clinical problem due to inhomogeneities within the diseased lung itself as well as the need to increase pressure or volume of oxygen to the lung as a life-saving measure. VILI is characterized by increased physical forces exerted within the lung, which results in cell death, inflammation and long-term fibrotic remodeling. Animal models can be used to study VILI, but it is challenging to distinguish the contributions of individual cell types in such a setup. In vitro models, which allow for controlled stretching of specific lung cell types have emerged as a potential option, but these models and the membranes used in them are unable to recapitulate some key features of the lung such as the 3D nanofibrous structure of the alveolar basement membrane while also allowing for cells to be cultured at an air liquid interface (ALI) and undergo increased mechanical stretch that mimics VILI. Here we develop a lung on a chip device with a nanofibrous synthetic membrane to provide ALI conditions and controllable stretching, including injurious stretching mimicking VILI. The lung on a chip device consists of a thin (i.e. ~20 μm) stretchable poly(caprolactone) (PCL) nanofibrous membrane placed between two channels fabricated in polydimethylsiloxane (PDMS) using 3D printed molds. We demonstrate that this lung on a chip device can be used to induce mechanotrauma in lung epithelial cells due to cyclic pathophysiologic stretch (~25%) that mimics clinical VILI. Pathophysiologic stretch induces cell injury and subsequently cell death, which results in loss of the epithelial monolayer, a feature mimicking the early stages of VILI. We also validate the potential of our lung on a chip device to be used to explore cellular pathways known to be altered with mechanical stretch and show that pathophysiologic stretch of lung epithelial cells causes nuclear translocation of the mechanotransducers YAP/TAZ. In conclusion, we show that a breathable lung on a chip device with a nanofibrous membrane can be easily fabricated using 3D printing of the lung on a chip molds and that this model can be used to explore pathomechanisms in mechanically induced lung injury.

Effect of interferon on Sindbis virus growth in chick-embryo cell cultures

1969 ◽  
Vol 15 (6) ◽  
pp. 605-610 ◽  
Author(s):  
Robert B. Stewart ◽  
Edward T. Sheaff
Keyword(s):  
Chick Embryo ◽  
Cell Cultures ◽  
Sindbis Virus ◽  
Embryo Cell ◽  
Virus Growth ◽  
Chick Embryo Cell ◽  
Embryo Cells ◽  
Number Of Cells

A study of the effect of interferon on the growth of Sindbis virus in cultures of chick-embryo cells has shown that interferon forms an association with cells (uptake) and that the action of interferon is concentration rather than amount dependent. Evidence has also been obtained that interferon acts to reduce the yield of virus from cells, but does not reduce the number of cells synthesizing virus (all or none effect).

scholarly journals Autophagy-associated Production of Antimicrobial Peptides hBD1 and LL37 Exhibits Anti-Bacillus Calmette-Guérin Effects in Lung Epithelial Cells

2020 ◽  
Author(s):  
Rui-ning Wang ◽  
Hong-lin Liu ◽  
Yao-xin Chen ◽  
Qian Wen ◽  
Xin-ying Zhou ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Antimicrobial Peptides ◽  
Active Form ◽  
Cell Types ◽  
Bactericidal Effect ◽  
Lung Epithelial Cells ◽  
Transcriptional Level ◽  
Lung Epithelial ◽  
Peptide Precursors ◽  
High Level

AbstractAntimicrobial peptides (AMPs) constitute important groups of bactericidal polypeptides against various microorganisms that exhibit their anti-bacteria activity through cleavage of precursor peptides into the active form of 50–100 amino acids in length. Various AMP cleavage mechanisms have been reported in different cell types; however, those in Mycobacterium tuberculosis (MTB)-infected lung epithelial cells remain unknown. In the present study, we found that MTB-infected lung epithelial cells expressed high level of the AMPs hBD1 and LL37 to kill intracellular MTB as the first-line immune barrier against MTB infection. Notably, their production in the lung epithelial cells was closely related to the function of autophagosomes and lysosomes. Experimental induction of autophagy in lung epithelial cells could enhance the expression of active hBD1 and LL37 at the post-transcriptional level, whereas silencing of these two active AMPs could decrease the bactericidal effect of autophagy. These findings indicated that cleavage of peptide precursors to form active AMPs might constitute a previously unrecognized antibacterial mechanism of autophagy.Author summaryLM and RW conceived and designed the experiments; RW performed the experiments and analyzed the data; QW analyzed the data and contributed reagents/materials/analysis tools; HL performed the experiments; XZ, JY, YL and ZH analyzed the data. LM and RW drafted the manuscript.

scholarly journals Shared epithelial pathways to lung repair and disease

2017 ◽  
Vol 26 (144) ◽  
pp. 170048 ◽  
Author(s):  
Magda Spella ◽  
Ioannis Lilis ◽  
Georgios T. Stathopoulos
Keyword(s):  
Epithelial Cells ◽  
Lung Diseases ◽  
Current Knowledge ◽  
Cell Types ◽  
Lung Epithelial Cells ◽  
Chronic Obstructive ◽  
Lung Repair ◽  
Chronic Lung Diseases ◽  
Lung Epithelial ◽  
Lung Regeneration

Chronic lung diseases present tremendous health burdens and share a common pathobiology of dysfunctional epithelial repair. Lung adenocarcinoma, the leading cancer killer worldwide, is caused mainly by chemical carcinogens of tobacco smoke that induce mutations in pulmonary epithelial cells leading to uncontrolled epithelial proliferation. Lung epithelial cells that possess the capacity for self-renewal and regeneration of other lung cell types are believed to underlie the pathobiology of chronic obstructive, fibrotic and neoplastic lung disorders. However, the understanding of lung epithelial progenitor cell hierarchy and turnover is incomplete and a comprehensive model of the cellular and transcriptional events that underlie lung regeneration and carcinogenesis is missing. The mapping of these processes is extremely important, since their modulation would potentially allow effective cure and/or prevention of chronic lung diseases. In this review we describe current knowledge on cellular and molecular pathways at play during lung repair and carcinogenesis and summarise the critical lung cell populations with regenerative and cancerous potential.

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.

Glycosaminoglycan expression in intestinal epithelial skin-fibroblastic cell cocultures. Fibroblastic cell-mediated effects of glucocorticoids

1989 ◽  
Vol 92 (4) ◽  
pp. 679-685
Author(s):  
F. Bouziges ◽  
P. Simon-Assmann ◽  
C. Leberquier ◽  
K. Haffen ◽  
M. Kedinger
Keyword(s):  
Molecular Weight ◽  
Cell Surface ◽  
Cell Cultures ◽  
Chondroitin Sulphate ◽  
Heparan Sulphate ◽  
Cell Types ◽  
Molecular Forms ◽  
Fibroblastic Cell ◽  
Endodermal Cells ◽  
Molecular Weight Form

The nature and distribution of newly synthesized glycosaminoglycans (GAGs) were studied in foetal rat skin fibroblasts, in rat intestinal endodermal cells and in cocultures of both cell types. The data show that fibroblasts synthesize and secrete hyaluronic acid (HA), heparan sulphate (HS) and chondroitin sulphate molecules (CS). Our data focus on HA, which is found as two different molecular forms, the smallest hydrodynamic-sized species being mostly recovered within the cell or associated with the cell surface, and the largest one secreted into the medium, whatever the cell type. Endodermal cells synthesize only two types of GAGs: the low molecular weight form of HA and HS. Cocultures of rat intestinal endodermal and skin fibroblastic cells in the presence of dexamethasone (Dx), allow optimal epithelial cytodifferentiation (Kedinger et al. 1987a). The main changes in the GAGs synthesized under these conditions as compared to skin fibroblastic cell cultures concern: (1) the enhancement of the lowest molecular weight form of HA to the detriment of the highest form in the cellular, pericellular and extracellular compartments; (2) the increase in the proportion of HS molecules associated with the cell surface. Interestingly, similar modifications are obtained by addition of Dx to the skin fibroblastic cell cultures. The data are discussed with reference to the constitution of a basement membrane at the epithelial-fibroblast interface in the cocultures, to the fibroblastic-dependent induction of epithelial differentiation and to the glucocorticoid response.

scholarly journals The effects of Con A on cell surface shedding in cell cultures

1980 ◽  
Vol 46 (1) ◽  
pp. 221-234
Author(s):  
T.C. Doetschman
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Muscle Cell ◽  
Cell Cultures ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Mouse Fibroblast ◽  
Specific Cell ◽  
Con A ◽  
A Cell

A cell surface immobilizing concentration of Con A inhibits the shedding of [3H]fucose-containing glycoproteins from the surface of chick embryonic leg and breast muscle cell cultures and cultures of the rat skeletal muscle cell line L6. The Con A-induced inhibition of shedding is much less in the mouse fibroblast cell line 3T3. In all 4 cell types the lectin inhibits the shedding of some fucosyl-glycoproteins more than others, especially those of a lipid-containing fraction which is excluded in Biogel A-5m chromatography. This differential nature of the Con A effect is not changed by the cytoskeletal disruptors cytochalasin B or colchicine. Con A causes an increase in the amount of trypsin-sensitive surface fucosylglycoprotein in the cell surface and appears to decrease the overall amount of cell surface degradation suggesting the inhibition of shedding caused by Con A is not due to an increase in internalization and degradation. The data suggest that some shedding may occur at specific cell surface sites to which surface materials must laterally migrate.

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