scholarly journals Micropatterning of Substrates for the Culture of Cell Networks by Stencil-Assisted Additive Nanofabrication

Micromachines ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 94
Author(s):  
Anita Previdi ◽  
Claudio Piazzoni ◽  
Francesca Borghi ◽  
Carsten Schulte ◽  
Leandro Lorenzelli ◽  
...  
Keyword(s):  
Cell Culture ◽  
Large Scale ◽  
Spatial Organization ◽  
Neuronal Cell ◽  
Morphological Properties ◽  
Patterned Films ◽  
Cluster Beam Deposition ◽  
Effective Additive ◽  
Cell Networks

The fabrication of in vitro neuronal cell networks where cells are chemically or electrically connected to form functional circuits with useful properties is of great interest. Standard cell culture substrates provide ensembles of cells that scarcely reproduce physiological structures since their spatial organization and connectivity cannot be controlled. Supersonic Cluster Beam Deposition (SCBD) has been used as an effective additive method for the large-scale fabrication of interfaces with extracellular matrix-mimicking surface nanotopography and reproducible morphological properties for cell culture. Due to the high collimation of SCBD, it is possible to exploit stencil masks for the fabrication of patterned films and reproduce features as small as tens of micrometers. Here, we present a protocol to fabricate micropatterned cell culture substrates based on the deposition of nanostructured cluster-assembled zirconia films by stencil-assisted SCBD. The effectiveness of this approach is demonstrated by the fabrication of micrometric patterns able to confine primary astrocytes. Calcium waves propagating in the astrocyte networks are shown.

LTCC Based Multi-Electrode Arrays for In-Vitro Cell Culture

2015 ◽  
Vol 2015 (CICMT) ◽  
pp. 000269-000274
Author(s):  
Heike Bartsch ◽  
Dirk Stöpel ◽  
Marcel Himmerlich ◽  
Martin Baca ◽  
Philipp Stadie ◽  
...  
Keyword(s):  
Cell Culture ◽  
Laser Scanning ◽  
Neuronal Cell ◽  
Laser Scanning Microscopy ◽  
Gold Electrodes ◽  
Neural Cells ◽  
Electrode Arrays ◽  
Green Tape ◽  
High Level

Neurobiological concepts based on state-of-the art technology have so far lacked the complexity of actual high-level neurobiological systems. Two key advances are needed to improve our understanding of such systems: in vitro 3D-neuronal cell culture and 3D MEA systems for measuring such 3D-cultures. These requirements call for smart multilayer and packaging technology. The material Green Tape TM from DuPont Nemours is chosen for the presented works, because its compatibility and those of available metallisation with cell cultures is already proven. An LTCC multilayer circuit with gold electrodes is the base of the 3D MEA. The layout of the 3D MEA is designed to fit the MEA2100-System for in vitro recording from Multi Channel Systems and enable thus a comparable data processing to established 2D MEAs Slots. The surface topography of the thick film electrodes and the surface state is investigated with laser scanning microscopy, SEM, XPS and measurements of the wetting angle of contact. The impedance of the screen printed electrodes is discussed taking these data into account. Their impedance amounts to 24 kΩ and are falls thus below the impedance of commercially available electroplated gold electrodes of 30 kΩ. First promising results have been achieved using 3D MEAs for 2D culture of human pluripotent stem cell derived neural cells.

scholarly journals Human Pluripotent Stem Cell Culture: Current Status, Challenges, and Advancement

2018 ◽  
Vol 2018 ◽  
pp. 1-17 ◽  
Author(s):  
Sushrut Dakhore ◽  
Bhavana Nayer ◽  
Kouichi Hasegawa
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Embryonic Stem ◽  
Current Status ◽  
Specific Cell ◽  
Therapeutic Applications ◽  
Proliferative Ability

Over the past two decades, human embryonic stem cells (hESCs) have gained attention due to their pluripotent and proliferative ability which enables production of almost all cell types in the human body in vitro and makes them an excellent tool to study human embryogenesis and disease, as well as for drug discovery and cell transplantation therapies. Discovery of human-induced pluripotent stem cells (hiPSCs) further expanded therapeutic applications of human pluripotent stem cells (PSCs). hPSCs provide a stable and unlimited original cell source for producing suitable cells and tissues for downstream applications. Therefore, engineering the environment in which these cells are grown, for stable and quality-controlled hPSC maintenance and production, is one of the key factors governing the success of these applications. hPSCs are maintained in a particular niche using specific cell culture components. Ideally, the culture should be free of xenobiotic components to render hPSCs suitable for therapeutic applications. Substantial efforts have been put to identify effective components, and develop culture conditions and protocols, for their large-scale expansion without compromising on quality. In this review, we discuss different media, their components and functions, including specific requirements to maintain the pluripotent and proliferative ability of hPSCs. Understanding the role of culture components would enable the development of appropriate conditions to promote large-scale, quality-controlled expansion of hPSCs thereby increasing their potential applications.

scholarly journals New ways to print living cells promise breakthroughs for engineering complex tissues in vitro

2006 ◽  
Vol 394 (2) ◽  
Author(s):  
Ginger S. Withers
Keyword(s):  
Tissue Engineering ◽  
Cell Growth ◽  
Cell Line ◽  
Neuronal Cell ◽  
Living Cells ◽  
Neuronal Cell Line ◽  
Biochemical Journal ◽  
Electrohydrodynamic Printing ◽  
Cell Networks

The ability to control the placement of cells and the assembly of networks in vitro has tremendous potential for understanding the regulation of development as well as for generating artificial tissues. To date, most engineering tools that can place materials with precision are not compatible with the requirements of living cells, and so approaches to tissue engineering have focused on patterning substrates as a way of controlling cell growth rather than patterning cells directly. In this issue of Biochemical Journal, however, Eagles et al. adapt electrohydrodynamic printing technology to ‘print’ living cells from a neuronal cell line on to a substrate. The importance of this approach is that it has the potential for unprecedented control over the position of cells in culture by directly placing them, thus allowing for the systematic assembly of cell networks.

scholarly journals Considerations for a Reliable In Vitro Model of Chemotherapy-Induced Peripheral Neuropathy

Toxics ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 300
Author(s):  
Sandy Eldridge ◽  
Arianna Scuteri ◽  
Eugenia M. C. Jones ◽  
Guido Cavaletti ◽  
Liang Guo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Peripheral Neuropathy ◽  
Human Cell ◽  
Anticancer Agents ◽  
In Vitro Model ◽  
Neuronal Cell ◽  
Severe Toxicity ◽  
Human Cell Culture ◽  
Vitro Model

Chemotherapy-induced peripheral neuropathy (CIPN) is widely recognized as a potentially severe toxicity that often leads to dose reduction or discontinuation of cancer treatment. Symptoms may persist despite discontinuation of chemotherapy and quality of life can be severely compromised. The clinical symptoms of CIPN, and the cellular and molecular targets involved in CIPN, are just as diverse as the wide variety of anticancer agents that cause peripheral neurotoxicity. There is an urgent need for extensive molecular and functional investigations aimed at understanding the mechanisms of CIPN. Furthermore, a reliable human cell culture system that recapitulates the diversity of neuronal modalities found in vivo and the pathophysiological changes that underlie CIPN would serve to advance the understanding of the pathogenesis of CIPN. The demonstration of experimental reproducibility in a human peripheral neuronal cell system will increase confidence that such an in vitro model is clinically useful, ultimately resulting in deeper exploration for the prevention and treatment of CIPN. Herein, we review current in vitro models with a focus on key characteristics and attributes desirable for an ideal human cell culture model relevant for CIPN investigations.

scholarly journals Advanced 3D Cell Culture Techniques in Micro-Bioreactors, Part I: A Systematic Analysis of the Literature Published between 2000 and 2020

Processes ◽  
2020 ◽  
Vol 8 (12) ◽  
pp. 1656
Author(s):  
Christoph Grün ◽  
Brigitte Altmann ◽  
Eric Gottwald
Keyword(s):  
Cell Culture ◽  
Large Scale ◽  
3D Cell Culture ◽  
Cell Types ◽  
Cell Contact ◽  
Systematic Analysis ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
3D Microenvironment

Bioreactors have proven useful for a vast amount of applications. Besides classical large-scale bioreactors and fermenters for prokaryotic and eukaryotic organisms, micro-bioreactors, as specialized bioreactor systems, have become an invaluable tool for mammalian 3D cell cultures. In this systematic review we analyze the literature in the field of eukaryotic 3D cell culture in micro-bioreactors within the last 20 years. For this, we define complexity levels with regard to the cellular 3D microenvironment concerning cell–matrix-contact, cell–cell-contact and the number of different cell types present at the same time. Moreover, we examine the data with regard to the micro-bioreactor design including mode of cell stimulation/nutrient supply and materials used for the micro-bioreactors, the corresponding 3D cell culture techniques and the related cellular microenvironment, the cell types and in vitro models used. As a data source we used the National Library of Medicine and analyzed the studies published from 2000 to 2020.

scholarly journals Neurophysiology of the Developing Cerebral Cortex: What We Have Learned and What We Need to Know

2022 ◽  
Vol 15 ◽  
Author(s):  
Heiko J. Luhmann
Keyword(s):  
Cerebral Cortex ◽  
Large Scale ◽  
Activity Patterns ◽  
Neuronal Cell ◽  
Electrode Arrays ◽  
Novel Technologies ◽  
The Past ◽  
Sensory Evoked

This review article aims to give a brief summary on the novel technologies, the challenges, our current understanding, and the open questions in the field of the neurophysiology of the developing cerebral cortex in rodents. In the past, in vitro electrophysiological and calcium imaging studies on single neurons provided important insights into the function of cellular and subcellular mechanism during early postnatal development. In the past decade, neuronal activity in large cortical networks was recorded in pre- and neonatal rodents in vivo by the use of novel high-density multi-electrode arrays and genetically encoded calcium indicators. These studies demonstrated a surprisingly rich repertoire of spontaneous cortical and subcortical activity patterns, which are currently not completely understood in their functional roles in early development and their impact on cortical maturation. Technological progress in targeted genetic manipulations, optogenetics, and chemogenetics now allow the experimental manipulation of specific neuronal cell types to elucidate the function of early (transient) cortical circuits and their role in the generation of spontaneous and sensory evoked cortical activity patterns. Large-scale interactions between different cortical areas and subcortical regions, characterization of developmental shifts from synchronized to desynchronized activity patterns, identification of transient circuits and hub neurons, role of electrical activity in the control of glial cell differentiation and function are future key tasks to gain further insights into the neurophysiology of the developing cerebral cortex.

scholarly journals Quantitative Study of Morphological Features of Stem Cells onto Photopatterned Azopolymer Films

2020 ◽  
Vol 11 (1) ◽  
pp. 8 ◽  
Author(s):  
Marcella Salvatore ◽  
Stefano Luigi Oscurato ◽  
Marietta D’Albore ◽  
Vincenzo Guarino ◽  
Stefania Zeppetelli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Large Scale ◽  
Human Mesenchymal Stem Cells ◽  
Cost Effective ◽  
Relevant Information ◽  
Material Transport ◽  
Optical Images ◽  
In Vitro Response ◽  
Patterned Films

In the last decade, the use of photolithography for the fabrication of structured substrates with controlled morphological patterns that are able to interact with cells at micrometric and nanometric size scales is strongly growing. A promising simple and versatile microfabrication method is based on the physical mass transport induced by visible light in photosensitive azobenzene-containing polymers (or azopolymers). Such light-driven material transport produces a modulation of the surface of the azopolymer film, whose geometry is controlled by the intensity and the polarization distributions of the irradiated light. Herein, two anisotropic structured azopolymer films have been used as substrates to evaluate the effects of topological signals on the in vitro response of human mesenchymal stem cells (hMSCs). The light-induced substrate patterns consist of parallel microgrooves, which are produced in a spatially confined or over large-scale areas of the samples, respectively. The analysis of confocal optical images of the in vitro hMSC cells grown on the patterned films offered relevant information about cell morphology—i.e., nuclei deformation and actin filaments elongation—in relation to the geometry and the spatial extent of the structured area of substrates. The results, together with the possibility of simple, versatile, and cost-effective light-induced structuration of azopolymers, promise the successful use of these materials as anisotropic platforms to study the cell guidance mechanisms governing in vitro tissue formation.

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