scholarly journals Electrophysiology Read-Out Tools for Brain-on-Chip Biotechnology

Micromachines ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 124
Author(s):  
Csaba Forro ◽  
Davide Caron ◽  
Gian Angotzi ◽  
Vincenzo Gallo ◽  
Luca Berdondini ◽  
...  
Keyword(s):  
Cell Biology ◽  
Brain Activity ◽  
Pharmaceutical Research ◽  
Brain Structures ◽  
Cell Culture Techniques ◽  
Lab On Chip ◽  
Culture Techniques ◽  
On Chip

Brain-on-Chip (BoC) biotechnology is emerging as a promising tool for biomedical and pharmaceutical research applied to the neurosciences. At the convergence between lab-on-chip and cell biology, BoC couples in vitro three-dimensional brain-like systems to an engineered microfluidics platform designed to provide an in vivo-like extrinsic microenvironment with the aim of replicating tissue- or organ-level physiological functions. BoC therefore offers the advantage of an in vitro reproduction of brain structures that is more faithful to the native correlate than what is obtained with conventional cell culture techniques. As brain function ultimately results in the generation of electrical signals, electrophysiology techniques are paramount for studying brain activity in health and disease. However, as BoC is still in its infancy, the availability of combined BoC–electrophysiology platforms is still limited. Here, we summarize the available biological substrates for BoC, starting with a historical perspective. We then describe the available tools enabling BoC electrophysiology studies, detailing their fabrication process and technical features, along with their advantages and limitations. We discuss the current and future applications of BoC electrophysiology, also expanding to complementary approaches. We conclude with an evaluation of the potential translational applications and prospective technology developments.

scholarly journals Organoids from the Human Fetal and Adult Pancreas

2019 ◽  
Vol 19 (12) ◽  
Author(s):  
Jeetindra R. A. Balak ◽  
Juri Juksar ◽  
Françoise Carlotti ◽  
Antonio Lo Nigro ◽  
Eelco J. P. de Koning
Keyword(s):  
Cell Biology ◽  
Cell Formation ◽  
Pancreatic Tissue ◽  
Ductal Adenocarcinoma ◽  
Disease Modelling ◽  
In Vivo Models ◽  
Pancreatic Cell ◽  
Culture Techniques

Abstract Purpose of Review Novel 3D organoid culture techniques have enabled long-term expansion of pancreatic tissue. This review comprehensively summarizes and evaluates the applications of primary tissue–derived pancreatic organoids in regenerative studies, disease modelling, and personalized medicine. Recent Findings Organoids derived from human fetal and adult pancreatic tissue have been used to study pancreas development and repair. Generated adult human pancreatic organoids harbor the capacity for clonal expansion and endocrine cell formation. In addition, organoids have been generated from human pancreatic ductal adenocarcinoma in order to study tumor behavior and assess drug responses. Summary Pancreatic organoids constitute an important translational bridge between in vitro and in vivo models, enhancing our understanding of pancreatic cell biology. Current applications for pancreatic organoid technology include studies on tissue regeneration, disease modelling, and drug screening.

scholarly journals Microtissues in Cardiovascular Medicine: Regenerative Potential Based on a 3D Microenvironment

2016 ◽  
Vol 2016 ◽  
pp. 1-20 ◽  
Author(s):  
Julia Günter ◽  
Petra Wolint ◽  
Annina Bopp ◽  
Julia Steiger ◽  
Elena Cambria ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Retention Rate ◽  
3D Cell Culture ◽  
Traditional Role ◽  
Cell Culture Techniques ◽  
Cell Therapies ◽  
Culture Techniques ◽  
3D Microenvironment

More people die annually from cardiovascular diseases than from any other cause. In particular, patients who suffer from myocardial infarction may be affected by ongoing adverse remodeling processes of the heart that may ultimately lead to heart failure. The introduction of stem and progenitor cell-based applications has raised substantial hope for reversing these processes and inducing cardiac regeneration. However, current stem cell therapies using single-cell suspensions have failed to demonstrate long-lasting efficacy due to the overall low retention rate after cell delivery to the myocardium. To overcome this obstacle, the concept of 3D cell culture techniques has been proposed to enhance therapeutic efficacy and cell engraftment based on the simulation of an in vivo-like microenvironment. Of great interest is the use of so-called microtissues or spheroids, which have evolved from their traditional role as in vitro models to their novel role as therapeutic agents. This review will provide an overview of the therapeutic potential of microtissues by addressing primarily cardiovascular regeneration. It will accentuate their advantages compared to other regenerative approaches and summarize the methods for generating clinically applicable microtissues. In addition, this review will illustrate the unique properties of the microenvironment within microtissues that makes them a promising next-generation therapeutic approach.

scholarly journals Organs-On-Chip Models of the Female Reproductive System

2019 ◽  
Vol 6 (4) ◽  
pp. 103 ◽  
Author(s):  
Vanessa Mancini ◽  
Virginia Pensabene
Keyword(s):  
Cell Biology ◽  
Reproductive Tract ◽  
Hormonal Treatment ◽  
Female Reproductive Tract ◽  
Specific Cell ◽  
Toxicity Screening ◽  
Paracrine Signalling ◽  
On Chip

Microfluidic-based technology attracts great interest in cell biology and medicine, in virtue of the ability to better mimic the in vivo cell microenvironment compared to conventional macroscale cell culture platforms. Recent Organs-on-chip (OoC) models allow to reproduce in vitro tissue and organ-level functions of living organs and systems. These models have been applied for the study of specific functions of the female reproductive tract, which is composed of several organs interconnected through intricate endocrine pathways and communication mechanisms. To date, a disease and toxicology study of this system has been difficult to perform. Thus, there is a compelling need to develop innovative platforms for the generation of disease model and for performing drug toxicity/screening in vitro studies. This review is focused on the analysis of recently published OoC models that recreate pathological and physiological characteristics of the female reproductive organs and tissues. These models aim to be used to assess changes in metabolic activity of the specific cell types and the effect of exposure to hormonal treatment or chemical substances on some aspects of reproduction and fertility. We examined these models in terms of device specifications, operating procedures, accuracy for studying the biochemical and functional activity of living tissues and the paracrine signalling that occurs within the different tissues. These models represent a powerful tool for understanding important diseases and syndromes affecting women all around the world. Immediate adoption of these models will allow to clarify diseases, causes and adverse events occurring during pregnancy such as pre-eclampsia, infertility or preterm birth, endometriosis and infertility.

scholarly journals Quantifying nanotherapeutics penetration using hydrogel based microsystem as a new 3D in-vitro platform

2021 ◽  
Author(s):  
Saba Goodarzi ◽  
Audrey Prunet ◽  
Fabien Rossetti ◽  
Olivier Tillement ◽  
Erika Porcel ◽  
...  
Keyword(s):  
Cell Biology ◽  
Practical Knowledge ◽  
Good Alternative ◽  
Intracellular Space ◽  
Tumour Spheroids ◽  
Specific Category ◽  
On Chip ◽  
Biology Laboratory

AbstractDespite progress in therapeutic strategies and understanding of cancer cell biology, there is a large attrition of promising therapeutics into the clinic. One predominant reason is the huge gap between 2D in-vitro assays used for drug screening, and the in-vivo 3D-physiological environment. This is particularly important for a specific category of emerging therapeutics: nanoparticles. The lack of physiological context hampered reliable predictions for the route and accumulation of those nanoparticles in-vivo. For such nanotherapeutics, Multi-Cellular Tumour Spheroids (MCTS) is emerging as a good alternative in-vitro model. However, the classical approaches to produce MCTS suffer from low yield, poor reproducibility and slow process, while spheroid-on-chip set-ups developed so far require a microfluidic practical knowledge difficult to transfer to a cell biology laboratory.We present here a simple yet highly flexible 3D-model microsystem consisting of agarose-based micro-wells. Fully compatible with the multi-well plates format conventionally used in cell biology, our simple process enables the formation of hundreds of reproducible spheroids in a single pipetting. It is compatible with live high-resolution optical microscopy and provides a user-friendly platform for in-situ immunostaining.As a proof-of-principle of the relevance of such in-vitro platform for the evaluation of nanoparticles, the aim of this study was to analyse the kinetic and localization of nanoparticles within colorectal cancer cells (HCT-116) MCTS. The nanoparticles chosen are sub-5 nm ultrasmall nanoparticles made of polysiloxane and gadolinium chelates that can be visualized in MRI and confocal microscopy (AGuIX®, currently implicated in clinical trials as effective radiosensitizers for radiotherapy). We show that the amount of AGuIX® nanoparticles within cells is largely different in 2D and 3D. Using our flexible agarose-based microsystems, we are able to resolve spatially and temporally the penetration and distribution of AGuIX® nanoparticles within tumour spheroids. The nanoparticles are first found in both extracellular and intracellular space of spheroids, within lysosomes compartment. While the extracellular part is washed away after few days, we evidenced trafficking of AGuIX® nanoparticles that are also found within mitochondria. Our agarose-based microsystem appears hence as a promising 3D in-vitro platform for investigation of nanotherapeutics transport, ahead of in-vivo studies.

scholarly journals A method to culture human alveolar rhabdomyosarcoma cell lines as rhabdospheres demonstrates an enrichment in stemness and notch signaling

Biology Open ◽  
2020 ◽  
pp. bio.050211
Author(s):  
Katherine K. Slemmons ◽  
Michael D. Deel ◽  
Yi-Tzu Lin ◽  
Kristianne M. Oristian ◽  
Nina Kuprasertkul ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Cell Lines ◽  
Three Dimensional ◽  
Notch Signaling Pathway ◽  
Stem Cell Markers ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Practical Tool

The development of three-dimensional cell culture techniques has allowed cancer researchers to study the stemness properties of cancer cells in in vitro culture. However, a method to grow PAX3-FOXO1 fusion-positive rhabdomyosarcoma (FP-RMS) - an aggressive soft tissue sarcoma of childhood - has to date not been reported, hampering efforts to identify the dysregulated signaling pathways that underlie FP-RMS stemness. Here, we first examine the expression of canonical stem cell markers in human RMS tumors and cell lines. We then describe a method to grow FP-RMS cell lines as rhabdospheres and demonstrate that these spheres are enriched in expression of canonical stemness factors as well as Notch signaling components. Specifically, FP-RMS rhabdospheres have increased expression of SOX2, POU5F1 (OCT4), and NANOG, and several receptors and transcriptional regulators in the Notch signaling pathway. FP-RMS rhabdospheres also exhibit functional stemness characteristics including multipotency, increased tumorigenicity in vivo, and chemoresistance. This method provides a novel practical tool to support research into FP-RMS stemness and chemoresistance signaling mechanisms.

scholarly journals An investigation into the possibility of bluetongue virus transmission by transfer of infected ovine embryos

2011 ◽  
Vol 78 (1) ◽  
Author(s):  
Estelle H. Venter ◽  
Truuske Gerdes ◽  
Isabel Wright ◽  
Johan Terblanche
Keyword(s):  
Cell Culture ◽  
Bluetongue Virus ◽  
Virus Transmission ◽  
Infected Sheep ◽  
Economic Losses ◽  
Bovine Embryos ◽  
Cell Culture Techniques ◽  
Culture Techniques

Bluetongue (BT), a disease that affects mainly sheep, causes economic losses owing to not only its deleterious effects on animals but also its associated impact on the restriction of movement of livestock and livestock germplasm. The causative agent, bluetongue virus (BTV), can occur in the semen of rams and bulls at the time of peak viraemia and be transferred to a developing foetus. The risk of the transmission of BTV by bovine embryos is negligible if the embryos are washed according to the International Embryo Transfer Society (IETS) protocol. Two experiments were undertaken to determine whether this holds for ovine embryos that had been exposed to BTV. Firstly, the oestrus cycles of 12 ewes were synchronised and the 59 embryos that were obtained were exposed in vitro to BTV-2 and BTV-4 at a dilution of 1 x 102.88 and 1 x 103.5 respectively. In the second experiment, embryos were recovered from sheep at the peak of viraemia. A total of 96 embryos were collected from BTV-infected sheep 21 days after infection. In both experiments half the embryos were washed and treated with trypsin according to the IETS protocol while the remaining embryos were neither washed nor treated. All were tested for the presence of BTV using cell culture techniques. The virus was detected after three passages in BHK-21 cells only in one wash bath in the first experiment and two unwashed embryos exposed to BTV-4 at a titre of 1 x 103.5. No embryos or uterine flush fluids obtained from viraemic donors used in the second experiment were positive for BTV after the standard washing procedure had been followed. The washing procedure of the IETS protocol can thus clear sheep embryos infected with BTV either in vitro or in vivo.

scholarly journals Applications of novel bioreactor technology to enhance the viability and function of cultured cells and tissues

2020 ◽  
Vol 10 (2) ◽  
pp. 20190090 ◽  
Author(s):  
H. W. Hoyle ◽  
L. A. Smith ◽  
R. J. Williams ◽  
S. A. Przyborski
Keyword(s):  
Cultured Cells ◽  
Ex Vivo ◽  
Low Cost ◽  
Tissue Slices ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Tissue Constructs ◽  
And Function

As the field of tissue engineering continues to advance rapidly, so too does the complexity of cell culture techniques used to generate in vitro tissue constructs, with the overall aim of mimicking the in vivo microenvironment. This complexity typically comes at a cost with regards to the size of the equipment required and associated expenses. We have developed a small, low-cost bioreactor system which overcomes some of the issues of typical bioreactor systems while retaining a suitable scale for the formation of complex tissues. Herein, we have tested this system with three cell populations/tissues: the culture of hepatocellular carcinoma cells, where an improved structure and basic metabolic function is seen; the culture of human pluripotent stem cells, in which the cultures can form more heterogeneous tissues resembling the in vivo teratoma and ex vivo liver tissue slices, in which improved maintenance of cellular viability is seen over the 3 days tested. This system has the flexibility to be used for a variety of further uses and has the potential to provide a more accessible alternative to current bioreactor technologies.

Cell Interactions With Polymers

2004 ◽  
Author(s):  
W. Mark Saltzman
Keyword(s):  
Cell Viability ◽  
Biomedical Applications ◽  
Polymer Surface ◽  
Polymeric Materials ◽  
Cell Interactions ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Wide Range

Synthetic and natural polymers are an important element in new strategies for producing engineered tissue. Polymers are currently used in a wide range of biomedical applications, including applications in which the polymer remains in intimate contact with cells and tissues for prolonged periods. As discussed in Chapter 1, several classes of polymers have proven to be most useful in biomedical applications and, therefore, might be appropriate for tissue engineering applications. To produce tissue-engineered materials composed of polymers and cells, however, it is first necessary to understand the influence of these polymeric materials on cell viability, growth, and function. Cell interactions with polymers are usually studied using cell culture techniques. While in vitro studies do not reproduce the wide range of cellular responses observed following implantation of materials, the culture environment provides a level of control and quantification that cannot usually be obtained in vivo. Cells in culture are generally plated over a polymer surface and the extent of cell adhesion and spreading on the surface can be measured. By maintaining the culture for longer periods the influence of the substrate on cell viability, function, and motility can also be determined. Since investigators use different techniques to assess cell interactions with polymers, and because the differences between techniques are critically important for interpretation of interactions, some of the most frequently used in vitro methods are reviewed in this section. Before any measurement of cell interaction with a polymer substrate can be attempted, the polymeric material and the cells must come into contact. Preferably, this contact should be controlled (or at least understood) by the experimentalist. This is a critical, and often overlooked, aspect of all of these measurements. Some materials are easily fabricated in a format suitable for study; polystyrene films, for example, are transparent, durable, and strong. Other materials must be coated onto a rigid substrate (such as a glass coverslip) prior to study. Cell function is sensitive to chemical, morphological, and mechanical properties of the surface; therefore, almost every aspect of material preparation can introduce variables that are known to influence cell interactions.

scholarly journals Gut-on-chip: Recreating human intestine in vitro

2020 ◽  
Vol 11 ◽  
pp. 204173142096531
Author(s):  
Yunqing Xiang ◽  
Hui Wen ◽  
Yue Yu ◽  
Mingqiang Li ◽  
Xiongfei Fu ◽  
...  
Keyword(s):  
Cell Biology ◽  
Intestinal Flora ◽  
In Vitro Models ◽  
Intestinal Microbiome ◽  
Human Gut ◽  
Intestinal Microbes ◽  
Number Of Microorganisms ◽  
On Chip

The human gut is important for food digestion and absorption, as well as a venue for a large number of microorganisms that coexist with the host. Although numerous in vitro models have been proposed to study intestinal pathology or interactions between intestinal microbes and host, they are far from recapitulating the real intestinal microenvironment in vivo. To assist researchers in further understanding gut physiology, the intestinal microbiome, and disease processes, a novel technology primarily based on microfluidics and cell biology, called “gut-on-chip,” was developed to simulate the structure, function, and microenvironment of the human gut. In this review, we first introduce various types of gut-on-chip systems, then highlight their applications in drug pharmacokinetics, host–gut microbiota crosstalk, and nutrition metabolism. Finally, we discuss challenges in this field and prospects for better understanding interactions between intestinal flora and human hosts, and then provide guidance for clinical treatment of related diseases.

scholarly journals Intestinal Organoids: New Tools to Comprehend the Virulence of Bacterial Foodborne Pathogens

Foods ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 108
Author(s):  
Mayra Aguirre Garcia ◽  
Killian Hillion ◽  
Jean-Michel Cappelier ◽  
Michel Neunlist ◽  
Maxime M. Mahe ◽  
...  
Keyword(s):  
Infectious Diseases ◽  
Pathogenic Bacteria ◽  
Foodborne Pathogen ◽  
Extensive Study ◽  
Infection Process ◽  
High Morbidity ◽  
Cell Culture Techniques ◽  
Culture Techniques

Foodborne diseases cause high morbidity and mortality worldwide. Understanding the relationships between bacteria and epithelial cells throughout the infection process is essential to setting up preventive and therapeutic solutions. The extensive study of their pathophysiology has mostly been performed on transformed cell cultures that do not fully mirror the complex cell populations, the in vivo architectures, and the genetic profiles of native tissues. Following advances in primary cell culture techniques, organoids have been developed. Such technological breakthroughs have opened a new path in the study of microbial infectious diseases, and thus opened onto new strategies to control foodborne hazards. This review sheds new light on cellular messages from the host–foodborne pathogen crosstalk during in vitro organoid infection by the foodborne pathogenic bacteria with the highest health burden. Finally, future perspectives and current challenges are discussed to provide a better understanding of the potential applications of organoids in the investigation of foodborne infectious diseases.

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