scholarly journals Cell behaviors within a confined adhesive area fabricated using novel micropatterning methods

PLoS ONE ◽  
2022 ◽  
Vol 17 (1) ◽  
pp. e0262632
Author(s):  
Tsukasa Nakatoh ◽  
Takuji Osaki ◽  
Sohma Tanimoto ◽  
Md. Golam Sarowar Jahan ◽  
Tomohisa Kawakami ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cell Division ◽  
Single Cells ◽  
Cell Behavior ◽  
Air Plasma ◽  
Cell Behaviors ◽  
Laser Ablation Method ◽  
Increasing Demand ◽  
Division Cell ◽  
Adhesive Coating

In the field of cell and tissue engineering, there is an increasing demand for techniques to spatially control the adhesion of cells to substrates of desired sizes and shapes. Here, we describe two novel methods for fabricating a substrate for adhesion of cells to a defined area. In the first method, the surface of the coverslip or plastic dish was coated with Lipidure, a non-adhesive coating material, and air plasma was applied through a mask with holes, to confer adhesiveness to the surface. In the second method, after the surface of the coverslip was coated with gold by sputtering and then with Lipidure; the Lipidure coat was locally removed using a novel scanning laser ablation method. These methods efficiently confined cells within the adhesive area and enabled us to follow individual cells for a longer duration, compared to the currently available commercial substrates. By following single cells within the confined area, we were able to observe several new aspects of cell behavior in terms of cell division, cell–cell collisions, and cell collision with the boundary between adhesive and non-adhesive areas.

ABOUT SOME PECULIARITIES OF THE PHYSIOLOGICAL HETEROGENEITY OF THE POPULATION OF SCENEDESMUS QUADRICAUDA (TURP.) BREB. IN THE PRESENCE OF LOW CONCENTRATIONS OF METALS

2018 ◽  
pp. 34-43
Author(s):  
V. I. Ipatova ◽  
A. G. Dmitrieva ◽  
О. F. Filenko ◽  
T. V. Drozdenko
Keyword(s):  
Cell Division ◽  
Toxic Effect ◽  
Copper Chloride ◽  
Single Cells ◽  
Physiological State ◽  
Scenedesmus Quadricauda ◽  
Physiological Status ◽  
Lag Phase ◽  
Protective Mechanism ◽  
Low Concentrations

The structure of the laboratory population of green microalgae Scenedesmus quadricauda (Turp.) Breb (=Desmodesmus communis E. Hegew.) was studied at different stages of its growth (lag-phase, log-phase and stationary phase) at low concentrations of copper chloride and silver nitrate by the method microculture, allowing to monitor the state and development of single cells having different physiological status. The response of the culture of S. quadricauda - the change in the number of cells and the fractional composition (the fraction of dividing, «dormant» and dying cells) depended not only on the concentration of the toxicant in the medium, but also on the physiological state of the culture: the level of synchronization and the growth phase. Silver ions at low concentrations had a more pronounced toxic effect on the culture than copper ions at different phases of its development, especially at a concentration of 0.001 mg/l (10-9 M). The main mechanism of the toxic effect of metals is to inhibit the process of cell division. At low concentrations of toxicants, especially at a concentration of 0.001 mg/l, a «paradoxical» effect expressed in the predominance of the fraction of «dormant» cells was revealed. The temporary inhibition of the process of cell division can be regarded as a protective mechanism that allows preserving the integrity of the population and its ability to survive in a changing environment. The obtained data explain the effect of action of low concentrations of substances due to their inclusion in the cell, the subsequent accumulation in the cell and their low excretion.

scholarly journals Extensive Regulation of Metabolism and Growth during the Cell Division Cycle

2014 ◽  
Author(s):  
Nikolai Slavov ◽  
David Botstein ◽  
Amy Caudy
Keyword(s):  
Gene Expression ◽  
Oxygen Consumption ◽  
Cell Division ◽  
Single Cell ◽  
Single Cells ◽  
Emergent Behavior ◽  
Yeast Cells ◽  
Physiological Data ◽  
Synchronized Cultures ◽  
Metabolic Cycle

Yeast cells grown in culture can spontaneously synchronize their respiration, metabolism, gene expression and cell division. Such metabolic oscillations in synchronized cultures reflect single-cell oscillations, but the relationship between the oscillations in single cells and synchronized cultures is poorly understood. To understand this relationship and the coordination between metabolism and cell division, we collected and analyzed DNA-content, gene-expression and physiological data, at hundreds of time-points, from cultures metabolically-synchronized at different growth rates, carbon sources and biomass densities. The data enabled us to extend and generalize our mechanistic model, based on ensemble average over phases (EAP), connecting the population-average gene-expression of asynchronous cultures to the gene-expression dynamics in the single-cells comprising the cultures. The extended model explains the carbon-source specific growth-rate responses of hundreds of genes. Our physiological data demonstrate that the frequency of metabolic cycling in synchronized cultures increases with the biomass density, suggesting that this cycling is an emergent behavior, resulting from the entraining of the single-cell metabolic cycle by a quorum-sensing mechanism, and thus underscoring the difference between metabolic cycling in single cells and in synchronized cultures. Measurements of constant levels of residual glucose across metabolically synchronized cultures indicate that storage carbohydrates are required to fuel not only the G1/S transition of the division cycle but also the metabolic cycle. Despite the large variation in profiled conditions and in the scale of their dynamics, most genes preserve invariant dynamics of coordination with each other and with the rate of oxygen consumption. Similarly, the G1/S transition always occurs at the beginning, middle or end of the high oxygen consumption phases, analogous to observations in human and drosophila cells. These results highlight evolutionary conserved coordination among metabolism, cell growth and division.

Patterns of cell division, cell death and chondrogenesis in cultured aggregates of normal and talpid3 mutant chick limb mesenchyme cells

Development ◽  
1972 ◽  
Vol 27 (1) ◽  
pp. 245-260
Author(s):  
D. A. Ede ◽  
O. P. Flint
Keyword(s):  
Cell Death ◽  
Cell Division ◽  
Limb Bud ◽  
Intercellular Matrix ◽  
Blue Stain ◽  
Special Pattern ◽  
Mesenchyme Cells ◽  
Division Cell ◽  
Made In

Aggregates were prepared from dissociated mesenchyme cells obtained from normal and talpid mutant chick limb buds at stage 26 and were maintained for 4 days in culture. They were shown by autoradiographic techniques to consist initially of populations of unifoimly dedifferentiated cells within which chondrogenesis was initiated between 1 and 2 days, leading to the formation of areas of precartilage in the interior of the aggregates. Measurements of cell population density, cell death and cell division were made in precartilage and non-cartilage regions on sections prepared from normal and mutant aggregates fixed at 1-day intervals and were related to the pattern of chondrogenesis. Non-cartilage areas consisted of cells surrounding the precartilage areas and extended to the surface of the aggregate; these cells showed no special pattern or histochemical reaction. Precartilage areas consisted of one or more “;condensations”, comprising cells arranged in concentric rings around a central cell or group of cells, characterized by uptake of [35S]sulphate and taking up alcian blue stain in the intercellular matrix. Chondrogenesis was initiated al the condensation foci and spread centrifugally. Condensations were arranged in a simple pattern, roughly equidistantly from each other and never at the surface of the aggregate. The shape and arrangement of the cells comprising them suggested that they were formed by a process of aggregation towards the condensation foci. The relation of these observations to events in the intact limb bud developing in vivo is discussed.

Anatomy of Wild Garlic Bulbs During and Subsequent to After-Ripening

Weed Science ◽  
1972 ◽  
Vol 20 (3) ◽  
pp. 233-237 ◽  
Author(s):  
J. F. Stritzke ◽  
E. J. Peters
Keyword(s):  
Cell Division ◽  
Microscopic Examination ◽  
Cell Elongation ◽  
Leaf Primordia ◽  
Root Primordia ◽  
Ripening Period ◽  
Shoot Primordia ◽  
Division Cell ◽  
Wild Garlic

Microscopic examination of central and soft offset bulbs of wild garlic(Allium vinealeL.) at senescence of the parent plants in May and June revealed embryonic plants with numerous root primordia and four or five shoot primordia. Hardshell bulbs and aerial bulblets contained only one or two root primordia and three leaf primordia. The embryonic plants of central, soft offset, and hardshell bulbs elongated slowly during the after-ripening period. Rapid cell division, cell elongation, and initiation of new leaves took place after termination of the after-ripening period in all but the dormant hardshell bulbs. In November, new hardshell bulbs could be seen at the base of plants developed from central and soft offset bulbs.

Electrospun cellulose-based conductive polymer nanofibrous mats: composite scaffolds and their influence on cell behavior with electrical stimulation for nerve tissue engineering

Soft Matter ◽  
2020 ◽  
Vol 16 (28) ◽  
pp. 6591-6598 ◽  
Author(s):  
Fangwen Zha ◽  
Wei Chen ◽  
Lu Hao ◽  
Chunsheng Wu ◽  
Meng Lu ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Electrical Stimulation ◽  
Nerve Cell ◽  
Conductive Polymer ◽  
Nerve Tissue ◽  
Cell Behavior ◽  
Composite Scaffolds ◽  
Nerve Tissue Engineering

Electrospun cellulose-based poly N-vinylpyrrole (PNVPY) and poly (3-hexylthiophene) (P3HT) nanofibrous mats and their influence on nerve cell behavior with electrical stimulation.

scholarly journals Digital Microfluidics for Single Bacteria Capture and Selective Retrieval Using Optical Tweezers

Micromachines ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 308 ◽  
Author(s):  
Phalguni Tewari Kumar ◽  
Deborah Decrop ◽  
Saba Safdar ◽  
Ioannis Passaris ◽  
Tadej Kokalj ◽  
...  
Keyword(s):  
Optical Tweezers ◽  
Cell Sorting ◽  
Magnetic Beads ◽  
Single Cells ◽  
Digital Microfluidics ◽  
Cell Behavior ◽  
Bacterial Cells ◽  
Microfluidic Platforms ◽  
Dynamic Cell ◽  
Downstream Analysis

When screening microbial populations or consortia for interesting cells, their selective retrieval for further study can be of great interest. To this end, traditional fluorescence activated cell sorting (FACS) and optical tweezers (OT) enabled methods have typically been used. However, the former, although allowing cell sorting, fails to track dynamic cell behavior, while the latter has been limited to complex channel-based microfluidic platforms. In this study, digital microfluidics (DMF) was integrated with OT for selective trapping, relocation, and further proliferation of single bacterial cells, while offering continuous imaging of cells to evaluate dynamic cell behavior. To enable this, magnetic beads coated with Salmonella Typhimurium-targeting antibodies were seeded in the microwell array of the DMF platform, and used to capture single cells of a fluorescent S. Typhimurium population. Next, OT were used to select a bead with a bacterium of interest, based on its fluorescent expression, and to relocate this bead to a different microwell on the same or different array. Using an agar patch affixed on top, the relocated bacterium was subsequently allowed to proliferate. Our OT-integrated DMF platform thus successfully enabled selective trapping, retrieval, relocation, and proliferation of bacteria of interest at single-cell level, thereby enabling their downstream analysis.

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