Contact Inhibition of Overlapping and Differential Cell Adhesion: A Sufficient Model for the Control of Certain Cell Culture Morphologies

1974 ◽  
Vol 16 (2) ◽  
pp. 401-419
Author(s):  
E. MARTZ ◽  
H. M. PHILLIPS ◽  
M. S. STEINBERG

Using intuitive arguments, several investigators have proposed that the relative strengths of adhesion of cell to cell and of cell to substratum could determine whether or not monolayering - and specifically contact inhibition of cell overlapping - will occur. In the present communication, these ‘strengths of adhesion’ are given precise physical definitions, and the adhesive relationships which would promote spontaneous cell monolayering are rigorously derived, using the thermodynamic approach embodied in the differential adhesion hypothesis. This analysis verifies that contact inhibition of overlapping could, in principle, be a result solely of differential adhesion. In addition, it is demonstrated that for homogeneous populations of uniform cells cultured on a solid, uniform substratum, eleven distinct equilibrium configurations (cell population morphologies) could be generated merely by varying the relative values of cell-to-cell and cell-to-substratum adhesiveness. Most of these configurations have been observed previously in actual cell cultures.

2000 ◽  
Vol 78 (3) ◽  
pp. 243-251 ◽  
Author(s):  
D A Beysens ◽  
G Forgacs ◽  
J A Glazier

Early embryonic development is characterized by spectacular morphogenetic processes such as sorting or spreading of tissues. Analogy between viscoelastic fluids and certain properties of embryonic tissues turned out to be useful in interpreting some aspects of these morphogenetic phenomena. In accordance with the differential adhesion hypothesis, the values of tissue-specific surface tensions have been shown to be consistent with the equilibrium configurations such tissues reach in the course of sorting and spreading. A method to measure tissue surface tension and viscoelastic properties is described. Notions like the Laplace's equation relating surface tension to radii of curvature, or the Kelvin model of viscoelasticity are used to analyze the results of these measurements. The fluid analogy is extended to time-dependent phenomena, in particular, to the analysis of cellular pattern evolution in the course of spreading. On the basis of recent experimental findings, we demonstrate that the kinetics of spreading and nucleation in binary fluids can be analyzed using the same formalism. We illustrate how our results can be used to obtain biologically relevant information on the strength of binding between specific cell adhesion molecules under near physiological conditions. We also suggest a diagnostic application of our method to monitor the metastatic potential of tumors. PACS No.: 03.65Ge


2017 ◽  
Vol 14 (132) ◽  
pp. 20170338 ◽  
Author(s):  
Harriet B. Taylor ◽  
Anaïs Khuong ◽  
Zhonglin Wu ◽  
Qiling Xu ◽  
Rosalind Morley ◽  
...  

Eph receptor and ephrin signalling has a major role in cell segregation and border formation, and may act through regulation of cell adhesion, repulsion or tension. To elucidate roles of cell repulsion and adhesion, we combined experiments in cell culture assays with quantitations of cell behaviour which are used in computer simulations. Cells expressing EphB2, or kinase-inactive EphB2 (kiEphB2), segregate and form a sharp border with ephrinB1-expressing cells, and this is disrupted by knockdown of N-cadherin. Measurements of contact inhibition of locomotion reveal that EphB2-, kiEphB2- and ephrinB1-expressing cells have strong heterotypic and weak homotypic repulsion. EphB2 cells have a transient increase in migration after heterotypic activation, which underlies a shift in the EphB2–ephrinB1 border but is not required for segregation or border sharpening. Simulations with the measured values of cell behaviour reveal that heterotypic repulsion can account for cell segregation and border sharpening, and is more efficient than decreased heterotypic adhesion. By suppressing homotypic repulsion, N-cadherin creates a sufficient difference between heterotypic and homotypic repulsion, and enables homotypic cohesion, both of which are required to sharpen borders.


2018 ◽  
Author(s):  
Tisong Liang ◽  
Rongfa Guan ◽  
Guozhou Cao ◽  
Haitao Shen ◽  
Zhenfeng Liu ◽  
...  

ABSTRACTThe 2D cell culture is the predominant in vitro model for numerous studies. However, 2D cell cultures may not accurately reflect the functions of three-dimensional (3D) tissues, which have extensive cell–cell and cell–matrix interactions; thus, using 2D cell cultures may lead to inaccurate experimental results. Therefore, to obtain adequate and detailed information about the antioxidant activity of cyanidin-3-O-glucoside (C3G) and C3G liposomes in the 2D and 3D cell culture models, we used in this study H2O2to construct the cell damage model and assess the antioxidant activity of C3G and C3G liposomes on Caco-2 cells cultured in the 3D model. We also measured the cell viability, cell morphology, and activity of glutathione (GSH), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) content of Caco-2 cells treated with H2O2, C3G, and C3G liposomes. Results showed that cells cultured in the 3D culture model formed a 3D structure and tight spheroids and showed increased cell activity and IC50. The C3G and C3G liposomes can enhance the activity of GSH, SOD, and T-AOC but decrease the MDA content. At the same time, the effect was more obvious in the 3D cell culture model than in the cells cultured in the 2D model. This study revealed that the results obtained from the 2D cell model may be inaccurate compared with the results obtained from the 3D cell model. A realistic mechanism study of antioxidant activity of C3G and C3G liposomes in the 3D cell model, which acts as an intermediate stage bridging the in vitro 2D and in vivo models, was observed.


1981 ◽  
Vol 49 (1) ◽  
pp. 217-223
Author(s):  
L.L. Wiseman ◽  
J. Strickler

Differential cell adhesion, a suggested guiding force for tissue rearrangement during embryogenesis, could be affected by desmosome frequency. A model system for studying embryonic tissue-positioning behaviour involves combining different tissues and following their rearrangements. We have previously shown that for one tissue, embryonic chick heart ventricle, direction of tissue positioning can be altered experimentally. Heart tissue precultured for 2.5 days tends to segregate internally, while tissue pre-cultured for just half a day tends to segregate externally. Also, intact fragments of tissue tend to segregate internally, while reaggregates of trypsin-disaggregated tissues tend to segregate externally. We show here that treatments that increase the tendency to internalize also increase the frequency of adherens junctions and treatments that increase the tendency to externalize decrease the frequency of junctions. An identical hierarchical ordering of the 4 experimental tissues occurs with respect to positioning behaviour and desmosome frequency. In the hierarchy, 2.5-day-cultured fragments greater than 2.5-day-cultured reaggregates greater than 0.5-day-cultured fragments 0.5-day-cultured reaggregates, tissues to the left tend to segregate internally and to have more desmosomes. Tissues to the right segregate externally and have fewer desmosome. This is what is expected if desmosome are organelles for adhesion and if differential adhesion is a factor in tissue-positioning behaviour.


Author(s):  
Andrzej Kociubiński ◽  
Dawid Zarzeczny ◽  
Maciej Szypulski ◽  
Aleksandra Wilczyńska ◽  
Dominika Pigoń ◽  
...  

The aim of the study was to present a method for assessing the condition of cell culture by measuring the impedance of cells cultured in the presence of nickel. For this purpose, an impedance measurement technique using nickel comb capacitors was used. The capacitor electrodes were made using a thin film magnetron sputtering. In the experimental part, the culture of cells of mouse fibroblasts on the prepared substrate was performed. The cell culture lasted 43 hours and showed that the presented technique allows it to be used to analyze the effect of nickel on cells.


2020 ◽  
Vol 36 (6) ◽  
pp. 35-48
Author(s):  
D.V. Коchkin ◽  
G.I. Sobolkovа ◽  
А.А. Fоmеnkov ◽  
R.А. Sidorov ◽  
А.М. Nоsоv

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.


2021 ◽  
Vol 22 (6) ◽  
pp. 3042
Author(s):  
Eun Ju Lee ◽  
Khurshid Ahmad ◽  
Shiva Pathak ◽  
SunJu Lee ◽  
Mohammad Hassan Baig ◽  
...  

In recent years, a major rise in the demand for biotherapeutic drugs has centered on enhancing the quality and efficacy of cell culture and developing new cell culture techniques. Here, we report fibronectin (FN) derived, novel peptides fibronectin-based intergrin binding peptide (FNIN)2 (18-mer) and FNIN3 (20-mer) which promote cell adhesion proliferation, and the differentiation of primary cells and stem cells. FNIN2 and 3 were designed based on the in silico interaction studies between FN and its receptors (integrin α5β1, αvβ3, and αIIbβ3). Analysis of the proliferation of seventeen-cell types showed that the effects of FNINs depend on their concentration and the existence of expressed integrins. Significant rhodamine-labeled FNIN2 fluorescence on the membranes of HeLa, HepG2, A498, and Du145 cells confirmed physical binding. Double coating with FNIN2 or 3 after polymerized dopamine (pDa) or polymerized tannic acid (pTA) precoating increased HBEpIC cell proliferation by 30–40 percent, suggesting FNINs potently affect primary cells. Furthermore, the proliferation of C2C12 myoblasts and human mesenchymal stem cells (MSCs) treated with FNINs was significantly increased in 2D/3D culture. FNINs also promoted MSC differentiation into osteoblasts. The results of this study offer a new approach to the production of core materials (e.g., cell culture medium components, scaffolds) for cell culture.


Author(s):  
Terry Riss ◽  
O. Joseph Trask

AbstractAlong with the increased use of more physiologically relevant three-dimensional cell culture models comes the responsibility of researchers to validate new assay methods that measure events in structures that are physically larger and more complex compared to monolayers of cells. It should not be assumed that assays designed using monolayers of cells will work for cells cultured as larger three-dimensional masses. The size and barriers for penetration of molecules through the layers of cells result in a different microenvironment for the cells in the outer layer compared to the center of three-dimensional structures. Diffusion rates for nutrients and oxygen may limit metabolic activity which is often measured as a marker for cell viability. For assays that lyse cells, the penetration of reagents to achieve uniform cell lysis must be considered. For live cell fluorescent imaging assays, the diffusion of fluorescent probes and penetration of photons of light for probe excitation and fluorescent emission must be considered. This review will provide an overview of factors to consider when implementing assays to interrogate three dimensional cell culture models.


1998 ◽  
Vol 10 (1) ◽  
pp. 3-10 ◽  
Author(s):  
G. M. Allan ◽  
F. McNeilly ◽  
S. Kennedy ◽  
B. Daft ◽  
J. A. Ellis ◽  
...  

Samples of lung, liver, kidney, pancreas, spleen, and lymph node from pigs with postweaning multisystemic wasting syndrome from California (USA) and samples of mesenteric lymph nodes from similarly diseased pigs from Brittany (France) were examined by light microscopy, in situ hybridization (ISH), and/or virus isolation. Whole genomic probes for porcine circovirus (PCV) and chicken anemia virus (CAV) were used for ISH. Tissue homogenate supernatants were inoculated onto PK/15 cells for virus isolation, and the presence of viral antigen and viral particles was verified by indirect immunofluorescence, ISH, and electron microscopy. Histologic examination of lung from pigs from California revealed interstitial pneumonia, alveolar epithelial hyperplasia, and basophilic nuclear and cytoplasmic inclusions in mononuclear cell infiltrates and various pulmonary epithelial cells. Granulomatous lymphadenitis with syncytial cells typified the lesions seen in the pigs from France. PCV-like nucleic acid was detected by ISH in lung, pancreas, lymph node, kidney, and liver in pigs from California. Positive signal was also obtained in lymph node sections from pigs from France. Probes for CAV were consistently negative. PK/15 cell cultures inoculated with lung preparations from diseased California pigs and mesenteric lymph node preparations from pigs from France had positive fluorescence by indirect staining for PCV using pooled polyclonal pig sera and hyperimmune rabbit serum and had variable staining with a panel of 7 monoclonal antibodies specific for cell culture contaminant PCV. PCV-like nucleic acid was also detected by ISH in cell cultures. Cytopathic effect was not observed Electron microscopic examination of inoculated cell cultures revealed 17-nm viral particles morphologically consistent with PCV No other virus particles were observed. Although genomic analysis for the definitive identification of these viral isolates remains to be done, the evidence provided strongly suggests that these tissue isolates are closely related to, although antigenically distinct from, the original PCV cell culture contaminant.


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