Cell movements in a confluent monolayer are not caused by gaps: evidence for direct contact inhibition of overlapping

1978 ◽  
Vol 30 (1) ◽  
pp. 293-304
Author(s):  
L. Timpe ◽  
E. Martz ◽  
M.S. Steinberg
Keyword(s):  
Direct Contact ◽  
Cell Monolayer ◽  
Cell Movement ◽  
Contact Inhibition ◽  
3T3 Cells ◽  
Confluent Monolayer ◽  
Cell Movements ◽  
Cell Locomotion ◽  
Confluent Cell ◽  
A Cell

According to the hypothesis of contact inhibition of movement, cells in a confluent monolayer are restrained from major overlapping by a directional inhibition of locomotion. This explanation of monolayering proposes that contact between 2 cells locally paralyses the locomotory function, preventing movement in the direction that would lead to overlapping. Consequently, a cell in contact on all sides with neighbouring cells should be immobilized. Yet in strictly monolayered cultures of confluent chick liver or mouse 3T3 cells, we have previously observed both translational cell movements and re-shufflings of relative cell positions. The ‘confluence’ was not perfect, however, and it seemed possible that the movements observed were due to release from contact inhibition by occasional transitory gaps seen to open up between cells. In the present study, detailed gap experiences and cell movements were recorded for 31 cells over a total of 1637 cell-hours. There was no significant correlation between frequency of gaps experienced and the extent of cell movement measured as neighbour-exchanges. We conclude that gaps are not a major cause of the movements observed. The hypothesis based on contact inhibition of motion, which attempts to explain monolayering indirectly by imposing a restraint on cell locomotion, cannot explain the substantial cell movements seen in the confluent cell monolayer studied here. To explain contact inhibition of overlapping, the evidence favours a more direct hypothesis which places no restriction on cell movement other than that overlapping be avoided. Such direct avoidance of overlapping could result from differences in the strengths with which cells adhere to one another and to the substratum.

scholarly journals Intracellular Position of Centrioles and the Direction of Homeostatic Epithelial Cell Movements in the Mouse Cornea

2016 ◽  
Vol 65 (2) ◽  
pp. 83-91 ◽  
Author(s):  
Erika Silverman ◽  
Jin Zhao ◽  
John C. Merriam ◽  
Takayuki Nagasaki
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Movement ◽  
Nerve Fibers ◽  
Narrow Strip ◽  
Cell Movements ◽  
Steady Rate ◽  
A Cell ◽  
Direction Of Movement ◽  
Basal Epithelial Cells

Corneal epithelial cells exhibit continuous centripetal movements at a rate of about 30 µm per day, but neither the driving force nor the mechanism that determines the direction of movements is known. To facilitate the investigation of homeostatic cell movement, we examined if the intracellular position of a centriole can be used as a directional marker of epithelial cell movements in the mouse cornea. A direction of cell movements was estimated in fixed specimens from a pattern of underlying subepithelial nerve fibers. Intracellular position of centrioles was determined by gamma-tubulin immunohistology and plotted in a narrow strip along the entire diameter of a cornea from limbus to limbus. When we determined the position of centrioles in the peripheral cornea where cell movements proceed generally along a radial path, about 55% of basal epithelial cells contained a centriole in the front half of a cell. However, in the central cornea where cells exhibit a spiral pattern of movements, centrioles were distributed randomly. These results suggest that centrioles tend to be positioned toward the direction of movement in corneal basal epithelial cells when they are moving centripetally at a steady rate.

Cell Crawling Assisted by Contractile Stress Induced Retraction

2010 ◽  
Vol 132 (6) ◽  
Author(s):  
Sitikantha Roy ◽  
Feng Miao ◽  
H. Jerry Qi
Keyword(s):  
Cell Movement ◽  
Focal Adhesions ◽  
Receptor Ligand ◽  
Produce Cell ◽  
Cell Locomotion ◽  
Cell Crawling ◽  
Parametric Studies ◽  
Contractile Stress ◽  
A Cell ◽  
Locomotion Speed

Cell locomotion is a result of a series of synchronized chemo-mechanical processes. Crawling-type cell locomotion consists of three steps: protrusion, translocation, and retraction. Previous works have shown that both protrusion and retraction can produce cell movement. For the latter, a cell derives its propulsive force from retraction induced protrusion mechanism, which was experimentally verified by Chen (1979, “Induction of Spreading During Fibroblast Movement,” J. Cell Biol., 81, pp. 684–691). In this paper, using finite element method, we take a computational biomimetic approach to study cell crawling assisted by contractile stress induced de-adhesion at the rear of the focal adhesion zone (FAZ). We assume the formation of the FAZ is driven by receptor-ligand bonds and nonspecific interactions. The contractile stress is generated due to the molecular activation of the intracellular actin-myosin machinery. The exerted contractile stress and its time dependency are modeled in a phenomenological manner as a two-spring mechanosensor proposed by Schwarz (2006, “Focal Adhesions as Mechanosensors: The Two-Spring Model,” BioSystems, 83(2–3), pp. 225–232). Through coupling the kinetics of receptor-ligand bonds with contractile stress, de-adhesion can be achieved when the stall value of the contractile stress is larger than a critical one. De-adhesion at the rear end of the FAZ causes a redistribution of elastic energy and induces cell locomotion. Parametric studies were conducted to investigate the connection between the cell locomotion speed and stall stress, and receptor-ligand kinetics. Finally, we provide a scaling relationship that can be used to estimate the cell locomotion speed.

scholarly journals Accurate Region-of-Interest Recovery Improves the Measurement of the Cell Migration Rate in the In Vitro Wound Healing Assay

2017 ◽  
Vol 22 (6) ◽  
pp. 626-635
Author(s):  
Cesar Bedoya ◽  
Andrés Cardona ◽  
July Galeano ◽  
Fabián Cortés-Mancera ◽  
Patrick Sandoz ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Monolayer ◽  
Region Of Interest ◽  
Wound Healing Assay ◽  
Time Intervals ◽  
Positioning Errors ◽  
Cellular Events ◽  
Confluent Cell ◽  
A Cell

The wound healing assay is widely used for the quantitative analysis of highly regulated cellular events. In this essay, a wound is voluntarily produced on a confluent cell monolayer, and then the rate of wound reduction (WR) is characterized by processing images of the same regions of interest (ROIs) recorded at different time intervals. In this method, sharp-image ROI recovery is indispensable to compensate for displacements of the cell cultures due either to the exploration of multiple sites of the same culture or to transfers from the microscope stage to a cell incubator. ROI recovery is usually done manually and, despite a low-magnification microscope objective is generally used (10x), repositioning imperfections constitute a major source of errors detrimental to the WR measurement accuracy. We address this ROI recovery issue by using pseudoperiodic patterns fixed onto the cell culture dishes, allowing the easy localization of ROIs and the accurate quantification of positioning errors. The method is applied to a tumor-derived cell line, and the WR rates are measured by means of two different image processing software. Sharp ROI recovery based on the proposed method is found to improve significantly the accuracy of the WR measurement and the positioning under the microscope.

Alterations of Ca2+ mobilizing properties in migrating endothelial cells

2001 ◽  
Vol 281 (2) ◽  
pp. H745-H754 ◽  
Author(s):  
Chiwaka Kimura ◽  
Masahiro Oike ◽  
Tetsuya Koyama ◽  
Yushi Ito
Keyword(s):  
Endothelial Cells ◽  
Cell Monolayer ◽  
Leading Edge ◽  
Cyclopiazonic Acid ◽  
Cellular Mechanisms ◽  
Intracellular Ca ◽  
Confluent Cell ◽  
A Cell ◽  
Endothelial Migration ◽  
Migrating Cells

Endothelial migration is one of the major events of pathological neovascularization. We compared the characteristics of Ca2+ mobilization in nonconfluent, confluent, and migrating endothelial cells. Migration of endothelial cells was induced by wounding the confluent cell monolayer. The basal intracellular Ca2+ concentration was lower in migrating cells and higher in confluent cells than in nonconfluent cells. Thapsigargin (TG)-induced Ca2+ leak and TG-evoked Ca2+ entry were accelerated in migrating cells, whereas the latter was suppressed in confluent cells. The ATP-induced Ca2+ transient was also much larger in migrating cells than in confluent cells. These alterations were also observed in a cell as an intracellular polarization, i.e., the leading edge showed an acceleration of TG-evoked Ca2+ entry and an augmentation of the ATP-induced Ca2+ transient. Endothelial migration was significantly suppressed by TG or cyclopiazonic acid. These observations suggest that the alterations of Ca2+ store site-related Ca2+ mobilizations, i.e., Ca2+sequestration, release, and TG-evoked Ca2+ entry, may be involved in the cellular mechanisms of endothelial migration.

Cell locomotion within a contact-inhibited monolayer of chick embryonic liver parenchyma cells

1975 ◽  
Vol 18 (3) ◽  
pp. 405-425
Author(s):  
D.R. Garrod ◽  
M.S. Steinberg
Keyword(s):  
Cell Movement ◽  
Liver Parenchyma ◽  
Time Lapse ◽  
Contact Inhibition ◽  
Embryonic Liver ◽  
Relative Movement ◽  
Cell Locomotion ◽  
Epitheloid Cells ◽  
Parenchyma Cells ◽  
Better Than

Using time-lapse filming, the relative movement of cells (nuclei) within a contact-inhibited monolayer of chick embryonic liver parenchyma cells has been studied. Two techniques were employed to determine the amount of relative cell movement during a culture period of 6 h. Firstly, the number of neighbours lost or gained by each nucleus was counted. Secondly, the relative distance moved by each nucleus in relation to other nucleus in the monolayer was measured. (The numerical results obtained from these analyses and details of the methods used are given in the text). A considerable amount of relative movement of nuclei within the monolayer was found during this period of culture. Although some gaps were occasionally seen between the cells in the monolayer, it was observed that cells able both to “ruffle” and to translocate when no gap was detectable; i.e. the cells appeared able to move while entirely surrounded by other cells. Because of this, we suggest that the monolayering of these epitheloid cells on a surface may be due to restriction of overlapping between them rather than to inhibition of movement by mutual contact. We argue that the term “contact inhibition of overlapping” relates to this behaviour better than the term “contact inhibition of movement”.

Analysis of the Dynamics of the Electric Capacitance of a Cell Monolayer at the Late Stages of Caco-2 cell Differentiation

2019 ◽  
Vol 35 (6) ◽  
pp. 87-90
Author(s):  
S.V. Nikulin ◽  
V.A. Petrov ◽  
D.A. Sakharov
Keyword(s):  
Impedance Spectroscopy ◽  
Cell Monolayer ◽  
Microfluidic Chips ◽  
Russian Science ◽  
Electric Capacitance ◽  
A Cell ◽  
Static Conditions ◽  
Late Stages

The real-time monitoring of electric capacitance (impedance spectroscopy) allowed obtaining evidence that structures which look like intestinal villi can be formed during the cultivation under static conditions as well as during the cultivation in microfluidic chips. It was shown in this work via transcriptome analysis that the Hh signaling pathway is involved in the formation of villus-like structures in vitro, which was previously shown for their formation in vivo. impedance spectroscopy, intestine, villi, electric capacitance, Hh The study was funded by the Russian Science Foundation (Project 16-19-10597).

Manipulating the Barrier Function of a Cell Monolayer Using a High-power Miniature Ultrasonic Transducer

2021 ◽  
Author(s):  
Mihnea V. Turcanu ◽  
Alexandru C. Moldovan ◽  
Maya Thanou ◽  
Inke Nathke ◽  
Sandy Cochran
Keyword(s):  
High Power ◽  
Barrier Function ◽  
Ultrasonic Transducer ◽  
Cell Monolayer ◽  
A Cell

Control of proliferin gene expression in serum-stimulated mouse cells

1987 ◽  
Vol 7 (6) ◽  
pp. 2080-2086
Author(s):  
D I Linzer ◽  
E L Wilder
Keyword(s):  
Protein Synthesis ◽  
S Phase ◽  
Mrna Levels ◽  
3T3 Cells ◽  
Serum Stimulation ◽  
Posttranscriptional Processing ◽  
A Cell ◽  
Mouse Cells ◽  
Stimulation Of

The serum-inducible expression of proliferin genes in BALB/c 3T3 cells was found to be dependent on both protein synthesis and an extended presence of serum in the medium. Even though no mature proliferin mRNA was detected in serum-starved cells, transcription of the proliferin genes occurred in these resting-cell cultures, indicating that posttranscriptional events may be important for regulating proliferin mRNA levels. These results suggest that protein synthesis after serum stimulation of quiescent mouse fibroblasts is required for posttranscriptional processing or stabilization of proliferin RNA. Proliferin RNA levels were found to be heterogeneous among serum-stimulated cells analyzed by in situ hybridization. This heterogeneity is probably due to asynchrony in the population and may point to a correlation between the time of proliferin expression and the time of entry of a cell into S phase.

scholarly journals 3D Bioprinted GelMA Based Models for the Study of Trophoblast Cell Invasion

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Houzhu Ding ◽  
Nicholas P. Illsley ◽  
Robert C. Chang
Keyword(s):  
Cell Invasion ◽  
Functional Limitations ◽  
Cell Movement ◽  
Trophoblast Cell ◽  
Trophoblast Invasion ◽  
Ring Model ◽  
Cell Front ◽  
A Cell ◽  
Practical Tool ◽  
And Migration

AbstractBioprinting is an emerging and promising technique for fabricating 3D cell-laden constructs for various biomedical applications. In this paper, we employed 3D bioprinted GelMA-based models to investigate the trophoblast cell invasion phenomenon, enabling studies of key placental functions. Initially, a set of optimized material and process parameters including GelMA concentration, UV crosslinking time and printing configuration were identified by systematic, parametric study. Following this, a multiple-ring model (2D multi-ring model) was tested with the HTR-8/SVneo trophoblast cell line to measure cell movement under the influence of EGF (chemoattractant) gradients. In the multi-ring model, the cell front used as a cell invasion indicator moves at a rate of 85 ± 33 µm/day with an EGF gradient of 16 µM. However, the rate was dramatically reduced to 13 ± 5 µm/day, when the multi-ring model was covered with a GelMA layer to constrain cells within the 3D environment (3D multi-ring model). Due to the geometric and the functional limitations of multi-ring model, a multi-strip model (2D multi-strip model) was developed to investigate cell movement in the presence and absence of the EGF chemoattractant. The results show that in the absence of an overlying cell-free layer of GelMA, movement of the cell front shows no significant differences between control and EGF-stimulated rates, due to the combination of migration and proliferation at high cell density (6 × 106 cells/ml) near the GelMA surface. When the model was covered by a layer of GelMA (3D multi-strip model) and migration was excluded, EGF-stimulated cells showed an invasion rate of 21 ± 3 µm/day compared to the rate for unstimulated cells, of 5 ± 4 µm/day. The novel features described in this report advance the use of the 3D bioprinted placental model as a practical tool for not only measurement of trophoblast invasion but also the interaction of invading cells with other tissue elements.

Sign in / Sign up

Export Citation Format

Share Document