scholarly journals Telocytes: ultrastructural, immunohistochemical and electrophysiological characteristics in human myometrium

Reproduction ◽  
2013 ◽  
Vol 145 (4) ◽  
pp. 357-370 ◽  
Author(s):  
Sanda M Cretoiu ◽  
Dragos Cretoiu ◽  
Adela Marin ◽  
Beatrice Mihaela Radu ◽  
Laurentiu M Popescu
Keyword(s):  
Contractile Activity ◽  
Three Dimensional ◽  
Time Lapse ◽  
Human Myometrium ◽  
Dynamic Interactions ◽  
Distinct Cell Type ◽  
Dimensional Network ◽  
Distinct Cell ◽  
Pregnant State

Telocytes (TCs) have been described in various organs and species (www.telocytes.com) as cells with telopodes (Tps) – very long cellular extensions with an alternation of thin segments (podomers) and dilated portions (podoms). We examined TCs using electron microscopy (EM), immunohistochemistry (IHC), immunofluorescence (IF), time-lapse videomicroscopy and whole-cell patch voltage clamp. EM showed a three-dimensional network of dichotomous-branching Tps, a labyrinthine system with homocellular and heterocellular junctions. Tps release extracellular vesicles (mean diameter of 160.6±6.9 nm in non-pregnant myometrium and 171.6±4.6 nm in pregnant myometrium), sending macromolecular signals to neighbouring cells. Comparative measurements (non-pregnant and pregnant myometrium) of podomer thickness revealed values of 81.94±1.77 vs 75.53±1.81 nm, while the podoms' diameters were 268.6±8.27 vs 316.38±17.56 nm. IHC as well as IF revealed double c-kit and CD34 positive results. Time-lapse videomicroscopy of cell culture showed dynamic interactions between Tps and myocytes. In non-pregnant myometrium, patch-clamp recordings of TCs revealed a hyperpolarisation-activated chloride inward current with calcium dependence and the absence of L-type calcium channels. TCs seem to have no excitable properties similar to the surrounding smooth muscle cells (SMCs). In conclusion, this study shows the presence of TCs as a distinct cell type in human non-pregnant and pregnant myometrium and describes morphometric differences between the two physiological states. In addition, we provide a preliminaryin vitroelectrophysiological evaluation of the non-pregnant state, suggesting that TCs could influence timing of the contractile activity of SMCs.

Tough Double-Network Hydrogels as Scaffolds for Tissue Engineering

2012 ◽  
pp. 213-222
Author(s):  
Jing Jing Yang ◽  
Jian Fang Liu ◽  
Takayuki Kurokawa ◽  
Nobuto Kitamura ◽  
Kazunori Yasuda ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Three Dimensional ◽  
Cartilage Regeneration ◽  
Cell Types ◽  
Embryoid Bodies ◽  
Living Tissue ◽  
Double Network ◽  
Dimensional Network

Hydrogels are used as scaffolds for tissue engineering in vitro & in vivo because their three-dimensional network structure and viscoelasticity are similar to those of the macromolecular-based extracellular matrix (ECM) in living tissue. Especially, the synthetic hydrogels with controllable and reproducible properties were used as scaffolds to study the behaviors of cells in vitro and implanted test in vivo. In this review, two different structurally designed hydrogels, single-network (SN) hydrogels and double-network (DN) hydrogels, were used as scaffolds. The behavior of two cell types, anchorage-dependent cells and anchorage-independent cells, and the differentiation behaviors of embryoid bodies (EBs) were investigated on these hydrogels. Furthermore, the behavior of chondrocytes on DN hydrogels in vitro and the spontaneous cartilage regeneration induced by DN hydrogels in vivo was examined.

scholarly journals The mechanism of MinD stability modulation by MinE in Min protein dynamics

2021 ◽  
Author(s):  
William C Carlquist ◽  
Eric N Cytrynbaum
Keyword(s):  
Time Course ◽  
Time Lapse ◽  
Membrane Stability ◽  
The Novel ◽  
Dynamic Interactions ◽  
Asymmetric Activation ◽  
Novel Model

The patterns formed both in vivo and in vitro by the Min protein system have attracted much interest because of the complexity of their dynamic interactions given the apparent simplicity of the component parts. Despite both the experimental and theoretical attention paid to this system, the details of the biochemical interactions of MinD and MinE, the proteins responsible for the patterning, are still unclear. For example, no model consistent with the known biochemistry has yet accounted for the observed dual role of MinE in the membrane stability of MinD. Until now, a statistical comparison of models to the time course of Min protein concentrations on the membrane has not been carried out. Such an approach is a powerful way to test existing and novel models that are difficult to test using a purely experimental approach. Here, we extract time series from previously published fluorescence microscopy time lapse images of in vitro experiments and fit two previously described and one novel mathematical model to the data. We find that the novel model, which we call the Asymmetric Activation with Bridged Stability Model, fits the time-course data best. It is also consistent with known biochemistry and explains the dual MinE role via MinE-dependent membrane stability that transitions under the influence of rising MinE to membrane instability with positive feedback. Our results reveal a more complex network of interactions between MinD and MinE underlying Min-system dynamics than previously considered.

scholarly journals Physical cues of biomaterials guide stem cell fate of differentiation: The effect of elasticity of cell culture biomaterials

Open Physics ◽  
2018 ◽  
Vol 16 (1) ◽  
pp. 943-955 ◽  
Author(s):  
Vincent Irawan ◽  
Akon Higuchi ◽  
Toshiyuki Ikoma
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Cell Fate ◽  
Empirical Model ◽  
Three Dimensional ◽  
Distinct Cell Type ◽  
Distinct Cell ◽  
Direct Sensing ◽  
D Cell ◽  
Culture Environment

Abstract It is widely accepted that stem cells directly sense the elasticity of two-dimensional (2-D) substrates and differentiate into a distinct cell type dependent on the substrate elasticity (direct-sensing differentiation: soft and hard substrates promote differentiation into soft and hard tissue lineage, respectively). Biologically, native extracellular matrices (ECMs) are constantly remodeled through out the life of individuals, which inadvertently introduce changes of mechanical properties. Therefore, direct-sensing differentiation might not fully take into account the responses of stem cells in the actual ECMs microenvironment. Recent investigations in three-dimensional (3-D) cell culture environment suggested the inconsistency of direct-sensing differentiation. Stem cells specifically differentiate not only by sensing the elasticity of materials but also by considering the cellular traction exerted to reorganize the matrices and the matrices deformation. This paper aims to expand further how the cells incorporate the elasticity cues and traction-mediated deformation in final differentiation fates of stem cells. To achieve the aim, we introduced an empirical model based on the investigations in 3-D cell culture environment. The empirical model would serve as a useful framework for future studies intended to investigate the relationship of traction-mediated deformation and commitment of stem cells for variety of tissue lineage in 2-D or 3-D cell culture environment.

scholarly journals Effect of Ethylenediaminetetraacetic Acid on Unsaturated Poly(Butylene Adipate-Co-Butylene Itaconate) Copolyester with Low-Melting Point and Controllable Hardness

Polymers ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 611 ◽  
Author(s):  
Chin-Wen Chen ◽  
Te-Sheng Hsu ◽  
Syang-Peng Rwei
Keyword(s):  
Melting Point ◽  
Textile Industry ◽  
Ethylenediaminetetraacetic Acid ◽  
Crosslinking Agent ◽  
Three Dimensional ◽  
Uv Curing ◽  
Plastic Properties ◽  
Smart Textile ◽  
Dimensional Network

A series of copolyesters, poly(butylene adipate-co-butylene itaconate) (PBABI), was synthesized using melt polycondensation from adipic acid (AA), itaconic acid (IA), 1,4-butanediol (1,4-BDO), and ethylenediaminetetraacetic acid (EDTA). 1H-NMR, FT-IR, GPC, DSC, TGA, DMA, XRD, Shore D, and tensile test were used to systematically characterize the structural and composition/physical properties of the copolyesters. It was found that the melting point (Tm) and crystallization temperature (Tc) of the copolyesters were, respectively, between 21.1 to 57.5 °C and −6.7 to 29.5 °C. The glass transition (Tg) and the initial thermal decomposition (Td-5%) temperatures of the PBABI copolyesters were observed to be between −53.6 to −55.8 °C and 313.6 and 342.1 °C at varying ratios of butylene adipate (BA) and butylene itaconate (IA), respectively. The XRD feature peak was identified at the 2θ values of 21.61°, 22.31°, and 23.96° for the crystal lattice of (110), (020), and (021), respectively. Interestingly, Shore D at various IA ratios had high values (between 51.3 to 62), which indicated that the PBABI had soft plastic properties. The Young’s modulus and elongation at break, at different IA concentrations, were measured to be at 0.77–128.65 MPa and 71.04–531.76%, respectively, which could be attributed to a close and compact three-dimensional network structure formed by EDTA as a crosslinking agent. There was a significant bell-shaped trend in a BA/BI ratio of 8/2, at different EDTA concentrations—the ∆Hm increased while the EDTA concentration increased from 0.001 to 0.05 mole% and then decreased at an EDTA ratio of 0.2 mole%. Since the PBABI copolymers have applications in the textile industry, these polymers have been adopted to reinforce 3D air-permeable polyester-based smart textile. This kind of composite not only possesses the advantage of lower weight and breathable properties for textiles, but also offers customizable, strong levels of hardness, after UV curing of the PBABI copolyesters, making its potential in vitro orthopedic support as the “plaster of the future”.

Evidence for the persistence of a decondensed chromosome scaffold in the interphase nucleus

1986 ◽  
Vol 86 (1) ◽  
pp. 155-171
Author(s):  
A.G. Bekers ◽  
A.C. Pieck ◽  
A.A. Rijken ◽  
F. Wanka
Keyword(s):  
Three Dimensional ◽  
Protein Composition ◽  
Bovine Liver ◽  
Dimensional Network ◽  
Dna Attachment ◽  
Cross Bridges ◽  
Triton X ◽  
Chromosome Scaffold

Nuclei of in vitro cultured bovine liver cells, deprived of the membranes by Triton X-100, were treated with 2 M-NaCl and DNase. Changes in ultrastructure and protein composition were studied at successive steps during treatment. Electron micrographs of nuclei treated with 2 M-NaCl showed a peripheral lamina and an internal system of randomly coiled filaments embedded in a mass of DNA fibres. After partial removal of the DNA the filaments could be seen to serve as backbones for the DNA attachment. Artificial redistribution occurring during fixation with glutaraldehyde suggests that the salt-resistant filaments are not stably cross-bridged into a three-dimensional network. The existence of reversible cross-bridges in vivo cannot be excluded, however. From the available data it is inferred that the filaments represent a decondensed from of the chromosome scaffolds and play a basic role in the organization of the genome throughout the nuclear cycle.

scholarly journals The lighthouse at the end of the chromosome*

F1000Research ◽  
2015 ◽  
Vol 4 ◽  
pp. 1427 ◽  
Author(s):  
Yahya Benslimane ◽  
Lea Harrington
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Spatial Organization ◽  
Three Dimensional ◽  
Time Lapse ◽  
Living Cells ◽  
Telomere Elongation ◽  
Cellular Processes ◽  
Time Lapse Microscopy

Fluorescence microscopy can be used to assess the dynamic localization and intensity of single entities in vitro or in living cells. It has been applied with aplomb to many different cellular processes and has significantly enlightened our understanding of the heterogeneity and complexity of biological systems. Recently, high-resolution fluorescence microscopy has been brought to bear on telomeres, leading to new insights into telomere spatial organization and accessibility, and into the mechanistic nuances of telomere elongation. We provide a snapshot of some of these recent advances with a focus on mammalian systems, and show how three-dimensional, time-lapse microscopy and single-molecule fluorescence shine a new light on the end of the chromosome.

scholarly journals RCCS Bioreactor-Based Modelled Microgravity Induces Significant Changes onIn Vitro3D Neuroglial Cell Cultures

2015 ◽  
Vol 2015 ◽  
pp. 1-14 ◽  
Author(s):  
Caterina Morabito ◽  
Nathalie Steimberg ◽  
Giovanna Mazzoleni ◽  
Simone Guarnieri ◽  
Giorgio Fanò-Illic ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Interactions ◽  
Microgravity Environment ◽  
Cell Aggregates ◽  
Differentiation Markers ◽  
Dynamic Interactions ◽  
Cx43 Expression ◽  
Wide Range ◽  
Cell Cell

We propose a human-derived neuro-/glial cell three-dimensionalin vitromodel to investigate the effects of microgravity on cell-cell interactions. A rotary cell-culture system (RCCS) bioreactor was used to generate a modelled microgravity environment, and morphofunctional features of glial-like GL15 and neuronal-like SH-SY5Y cells in three-dimensional individual cultures (monotypic aggregates) and cocultures (heterotypic aggregates) were analysed. Cell survival was maintained within all cell aggregates over 2 weeks of culture. Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation). In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool forin vitrostudies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.

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