Surfaces Designed to Control the Projected Area and Shape of Individual Cells

1999 ◽  
Vol 121 (1) ◽  
pp. 40-48 ◽  
Author(s):  
C. H. Thomas ◽  
J.-B. Lhoest ◽  
D. G. Castner ◽  
C. D. McFarland ◽  
K. E. Healy
Keyword(s):  
Mammalian Cells ◽  
Cell Function ◽  
Single Cells ◽  
Polymer Network ◽  
Interpenetrating Polymer Network ◽  
Specific Cell ◽  
Cell Binding ◽  
Projected Area ◽  
Cell Functions ◽  
Spatially Resolved

Materials with spatially resolved surface chemistry were designed to isolate individual mammalian cells to determine the influence of projected area on specific cell functions (e.g., proliferation, cytoskeletal organization). Surfaces were fabricated using a photolithographic process resulting in islands of cell binding N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (EDS) separated by a nonadhesive interpenetrating polymer network [poly (acrylamide-co-ethylene glycol); P(AAm-co-EG)]. The surfaces contained over 3800 adhesive islands/cm2, allowing for isolation of single cells with projected areas ranging from 100 μm2 to 10,000 μm2. These surfaces provide a useful tool for researching how cell morphology and mechanical forces affect cell function.

Materials Designed to Control and Examine The Function of Single Cells

1998 ◽  
Vol 530 ◽  
Author(s):  
C.H. Thomas ◽  
J.B. Lhoest ◽  
D.G. Castner ◽  
C.D. Mcfarland ◽  
K.E. Healy
Keyword(s):  
Mammalian Cells ◽  
Cell Function ◽  
Single Cells ◽  
Polymer Network ◽  
Interpenetrating Polymer Network ◽  
Cytoskeletal Organization ◽  
Spatially Resolved ◽  
Cell Organization ◽  
A Cell ◽  
And Function

AbstractStress levels imparted on a cell have been shown to alter cell organization and function, presumably as a result of morphological cues affecting cytoskeletal organization. Materials with spatially resolved surface chemistry were designed to isolate individual mammalian cells to determine the influence of projected area on cell proliferation and cytoskeletal organization. Surfaces were fabricated using a photolithographic process resulting in islands of cell binding N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (EDS) separated by a non-adhesive interpenetrating polymer network [poly acrylamide-co-ethylene glycol; P(AAm-co-EG)]. The surfaces contained over 3800 adhesive islands/cm2, allowing for isolation of single cells with projected areas ranging from 100µm2to 10,000µm2. These surfaces provide a useful tool for researching how cell morphology and mechanical forces affect cell function.

New views inside cells with a digital imaging microscope

1991 ◽  
Vol 49 ◽  
pp. 240-241
Author(s):  
F. S. Fay ◽  
Kevin Fogarty ◽  
Richard Tuft ◽  
Walter Carrington
Keyword(s):  
Digital Imaging ◽  
Cell Biology ◽  
Cell Function ◽  
Single Cells ◽  
Low Noise ◽  
Molecular Composition ◽  
Specific Cell ◽  
Wide Field ◽  
High Quantum Efficiency ◽  
Cell Functions

Many current questions in cell biology revolve around questions regarding how changes in cell function are caused by changes in their molecular composition. Given that cells are highly organized structures often carrying out diverse functions in different compartments, it follows that changes in specific cell functions must involve highly localized changes in molecular composition.We have been involved in the development of the digital imaging microscope as a tool to investigate the distribution of molecules inside single living cells. The system measures fluorescence of probes that are highly fluorescent and specific for a molecule or ion of interest and utilizes a wide-field rather than a confocal microscope to produce an image of fluorescence in a single cell. The image is captured by a high quantum efficiency, low noise cooled CCD, thereby providing ultrahigh efficiency in the acquisition of fluorescent images. By utilizing very powerful light sources, the system is capable of generating an image with good signal-to-noise ratio in a millisecond or less, thereby allowing one to follow extremely rapid changes in molecular or ion distribution in single cells.

scholarly journals Identification and further characterization of the specific cell binding fragment from sponge aggregation factor.

1986 ◽  
Vol 102 (4) ◽  
pp. 1344-1349 ◽  
Author(s):  
M Gramzow ◽  
M Bachmann ◽  
G Uhlenbruck ◽  
A Dorn ◽  
W E Müller
Keyword(s):  
Single Cells ◽  
Sodium Dodecyl ◽  
Cell System ◽  
Specific Cell ◽  
Cell Binding ◽  
Aggregation Factor ◽  
Indirect Immunofluorescence Staining ◽  
Number Of Binding Sites ◽  
Ca Ions

Monoclonal antibodies (McAbs) were raised against the aggregation factor (AF) from the marine sponge Geodia cydonium. Two clones were identified that secrete McAbs against the cell binding protein of the AF complex. Fab fragments of McAbs: 5D2-D11 completely abolished the activity of the AF to form secondary aggregates from single cells. The McAbs were determined to react with the AF in vitro; this interaction was prevented by addition of the aggregation receptor, isolated and purified from the same species. After dissociation of the AF by sodium dodecyl sulfate and 2-mercaptoethanol, followed by electrophoretical fractionation, a 47-kD protein was identified by immunoblotting which interacted with the McAbs: 5D2-D11. During this dissociation procedure, the sunburst structure of the AF was destroyed. In a second approach, the 47-kD protein was isolated by immunoprecipitation; 12 molecules of this protein species were calculated to be associated with the intact AF particle. The 47-kD AF fragment bound to dissociated Geodia cells with a high affinity (Ka of 7 X 10(8) M-1) even in the absence of Ca++ ions; the number of binding sites was approximately 4 X 10(6)/cell. This interaction was prevented by addition of the aggregation receptor to the 47-kD protein in the homologous cell system. Moreover, it was established that this binding occurs species-specifically. The 47-kD fragment of the AF was localized only extracellularly by indirect immunofluorescence staining in cryostat slices. These data suggest that the 47-kD protein is the cell binding molecule of the AF from Geodia.

scholarly journals A New Take on Prion Protein Dynamics in Cellular Trafficking

2020 ◽  
Vol 21 (20) ◽  
pp. 7763
Author(s):  
Rodrigo Nunes Alves ◽  
Rebeca Piatniczka Iglesia ◽  
Mariana Brandão Prado ◽  
Maria Isabel Melo Escobar ◽  
Jacqueline Marcia Boccacino ◽  
...  
Keyword(s):  
Prion Protein ◽  
Cell Function ◽  
Cellular Prion Protein ◽  
Specific Cell ◽  
Membrane Domains ◽  
Cellular Processes ◽  
Cell Functions ◽  
Heterogeneous Membrane ◽  
Cell Compartments ◽  
Distinct Cell

The mobility of cellular prion protein (PrPC) in specific cell membrane domains and among distinct cell compartments dictates its molecular interactions and directs its cell function. PrPC works in concert with several partners to organize signaling platforms implicated in various cellular processes. The scaffold property of PrPC is able to gather a molecular repertoire to create heterogeneous membrane domains that favor endocytic events. Dynamic trafficking of PrPC through multiple pathways, in a well-orchestrated mechanism of intra and extracellular vesicular transport, defines its functional plasticity, and also assists the conversion and spreading of its infectious isoform associated with neurodegenerative diseases. In this review, we highlight how PrPC traffics across intra- and extracellular compartments and the consequences of this dynamic transport in governing cell functions and contributing to prion disease pathogenesis.

scholarly journals Single-cell nucleic acid profiling in droplets (SNAPD) enables high-throughput analysis of heterogeneous cell populations

2020 ◽  
Author(s):  
Leland B. Hyman ◽  
Clare R. Christopher ◽  
Philip A. Romero
Keyword(s):  
Nucleic Acid ◽  
Single Cell ◽  
Mammalian Cells ◽  
Cancer Biology ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Cell Types ◽  
Cell Populations ◽  
Specific Cell ◽  
High Throughput Analysis

AbstractExperimental methods that capture the individual properties of single cells are revealing the key role of cell-to-cell variability in countless biological processes. These single-cell methods are becoming increasingly important across the life sciences in fields such as immunology, regenerative medicine, and cancer biology. Existing single-cell analysis methods are often limited by their low analysis throughput, their inability to profile high-dimensional phenotypes, and complicated experimental workflows with slow turnaround times. In this work, we present Single-cell Nucleic Acid Profiling in Droplets (SNAPD) to analyze the transcriptional states of hundreds of thousands of single mammalian cells. Individual cells are encapsulated in aqueous droplets on a microfluidic chip and the content of each cell is profiled by amplifying a targeted panel of transcriptional markers. Molecular logic circuits then integrate this multi-dimensional information to categorize cells based on their transcriptional profile and produce a detectable fluorescence output. SNAPD analyzes over 100,000 cells per hour and can be used to quantify distinct cell types within populations, detect rare cells at frequencies down to 0.1%, and enrich specific cell types using microfluidic sorting. SNAPD provides a simple, rapid, low cost, and scalable approach to study complex phenotypes in heterogeneous cell populations.

scholarly journals New Technologies for Use in Toxicology Studies: Monitoring the Effects of Xenobiotics on Immune Function

1990 ◽  
Vol 9 (3) ◽  
pp. 303-317 ◽  
Author(s):  
J. Paul Robinson ◽  
R.W. Pfeifer
Keyword(s):  
Flow Cytometry ◽  
New Technologies ◽  
Single Cells ◽  
Specific Cell ◽  
Kinetic Measurements ◽  
New Developments ◽  
Cell Functions ◽  
Alternative Approach ◽  
In Vitro Systems

New developments in flow cytometry are now being applied in toxicology studies. There are several reasons for using this technology. First, techniques are well characterized to measure functional parameters of single cells. Such measurements can be directly related to perturbations by xenobiotics, cell-mediated immune responses, or trauma. Second, there is a clear indication for movement toward in vitro systems as highly objective assessments of toxicologic interactions. By measuring specific cell functions at the single cell level, it is possible to define a range of normal responses. More importantly, a multiparametric analysis can be performed with flow cytometry and parameters can be directly related to one another. Furthermore, kinetic measurements can be made, providing vital clues to the mechanisms of actions of drugs or chemicals on functions of specific cell populations. Major advantages of this approach are that studies can be performed on very small volumes of blood, body fluid, or cell culture lines and it is not necessary to isolate pure populations of cells to perform these assays. We believe that this alternative approach in toxicology will provide valuable information unobtainable by traditional means.

Use of Flexible Materials with a Novel Pressure Driven Equibiaxial Cell Stretching Device for Mechanical Stimulation of Single Mammalian Cells

2003 ◽  
Vol 773 ◽  
Author(s):  
James D. Kubicek ◽  
Stephanie Brelsford ◽  
Philip R. LeDuc
Keyword(s):  
Cell Fate ◽  
Mechanical Stimulation ◽  
Mammalian Cells ◽  
Cellular Response ◽  
Single Cells ◽  
Complex Nature ◽  
Cellular Behavior ◽  
Cell Stretching ◽  
Stimulation Of ◽  
Pressure Driven

AbstractMechanical stimulation of single cells has been shown to affect cellular behavior from the molecular scale to ultimate cell fate including apoptosis and proliferation. In this, the ability to control the spatiotemporal application of force on cells through their extracellular matrix connections is critical to understand the cellular response of mechanotransduction. Here, we develop and utilize a novel pressure-driven equibiaxial cell stretching device (PECS) combined with an elastomeric material to control specifically the mechanical stimulation on single cells. Cells were cultured on silicone membranes coated with molecular matrices and then a uniform pressure was introduced to the opposite surface of the membrane to stretch single cells equibiaxially. This allowed us to apply mechanical deformation to investigate the complex nature of cell shape and structure. These results will enhance our knowledge of cellular and molecular function as well as provide insights into fields including biomechanics, tissue engineering, and drug discovery.

Metabolic Regulation and Related Molecular Mechanisms in Various Stem Cell Functions

2020 ◽  
Vol 15 (6) ◽  
pp. 531-546 ◽  
Author(s):  
Hwa-Yong Lee ◽  
In-Sun Hong
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Oxidative Phosphorylation ◽  
Metabolic Pathways ◽  
Critical Role ◽  
The Self ◽  
Specific Cell ◽  
Differentiation Potential ◽  
Self Renewal ◽  
Cell Functions

Recent studies on the mechanisms that link metabolic changes with stem cell fate have deepened our understanding of how specific metabolic pathways can regulate various stem cell functions during the development of an organism. Although it was originally thought to be merely a consequence of the specific cell state, metabolism is currently known to play a critical role in regulating the self-renewal capacity, differentiation potential, and quiescence of stem cells. Many studies in recent years have revealed that metabolic pathways regulate various stem cell behaviors (e.g., selfrenewal, migration, and differentiation) by modulating energy production through glycolysis or oxidative phosphorylation and by regulating the generation of metabolites, which can modulate multiple signaling pathways. Therefore, a more comprehensive understanding of stem cell metabolism could allow us to establish optimal culture conditions and differentiation methods that would increase stem cell expansion and function for cell-based therapies. However, little is known about how metabolic pathways regulate various stem cell functions. In this context, we review the current advances in metabolic research that have revealed functional roles for mitochondrial oxidative phosphorylation, anaerobic glycolysis, and oxidative stress during the self-renewal, differentiation and aging of various adult stem cell types. These approaches could provide novel strategies for the development of metabolic or pharmacological therapies to promote the regenerative potential of stem cells and subsequently promote their therapeutic utility.

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