scholarly journals A biomimetic liver model recapitulating bio-physical properties and tumour stroma interactions in hepatocellular carcinoma

2020 ◽  
Author(s):  
C. Calitz ◽  
N. Pavlovic ◽  
J. Rosenquist ◽  
C. Zagami ◽  
A. Samanta ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Culture ◽  
Liver Disease ◽  
Physical Properties ◽  
Chronic Liver Disease ◽  
3D Model ◽  
Stromal Compartment ◽  
Culture Models ◽  
Cell Culture Models

ABSTRACTHepatocellular carcinoma (HCC) is a primary liver tumor developing in the wake of chronic liver disease. Chronic liver disease and inflammation leads to a fibrotic environment actively supporting and driving hepatocarcinogenesis. Insight into hepatocarcinogenesis in terms of the interplay between the tumor stroma micro-environment and tumor cells is thus of considerable importance. Three-dimensional (3D) cell culture models are proposed as the missing link between current in vitro 2D cell culture models and in vivo animal models. Our aim was to design a novel 3D biomimetic HCC model with accompanying fibrotic stromal compartment and vasculature. Physiologically relevant hydrogels such as collagen and fibrinogen were incorporated to mimicking the bio-physical properties of the tumor ECM. In our model LX2 and HepG2 cells embedded in a hydrogel matrix were seeded onto the inverted insert membrane of a Transwell™ system. HUVEC cells were then seeded onto the opposite side of the membrane. Three formulations consisting of ECM-hydrogels embedded with cells were prepared and the bio-physical properties determined by rheology. Cell viability was determined by the AlamarBlue® assay over 21-days. The effect of the chemotherapeutic drug doxorubicin was evaluated in both a 2D co-culture and our 3D model for a period of 72h. We show that this model is viable for 25-days and gives rise to metastatic tumor nodules after 17 days in culture. Rheology results show that bio-physical properties of a fibrotic, cirrhotic and HCC liver can be successfully mimicked. Overall, results indicate that this 3D model is more representative of the in vivo situation compared to traditional 2D cultures. Our 3D tumor model showed a decreased response to chemotherapeutics, mimicking drug resistance typically seen in HCC patients. This model could in future provide a valuable new platform to study multifocal HCC or to identify mechanisms that contribute to early stages of metastasis.SUMMARYA protocol for a novel 3D biomimetic HCC model with accompanying fibrotic stromal compartment and vasculature, to study endocrine and paracrine signaling in liver cancer. The model uses physiological relevant hydrogels in ratios mimicking the bio-physical properties of the stromal extracellular matrix, which is an active mediator of cellular interactions, tumor growth and metastasis.

scholarly journals The Communication between Ocular Surface and Nasal Epithelia in 3D Cell Culture Technology for Translational Research: A Narrative Review

2021 ◽  
Vol 22 (23) ◽  
pp. 12994
Author(s):  
Malik Aydin ◽  
Jana Dietrich ◽  
Joana Witt ◽  
Maximiliane S. C. Finkbeiner ◽  
Jonas J.-H. Park ◽  
...  
Keyword(s):  
Cell Culture ◽  
Nasal Cavity ◽  
Ocular Surface ◽  
Measles Virus ◽  
3D Culture ◽  
3D Cell Culture ◽  
Narrative Review ◽  
Culture Models ◽  
Cell Culture Models

There is a lack of knowledge regarding the connection between the ocular and nasal epithelia. This narrative review focuses on conjunctival, corneal, ultrastructural corneal stroma, and nasal epithelia as well as an introduction into their interconnections. We describe in detail the morphology and physiology of the ocular surface, the nasolacrimal ducts, and the nasal cavity. This knowledge provides a basis for functional studies and the development of relevant cell culture models that can be used to investigate the pathogenesis of diseases related to these complex structures. Moreover, we also provide a state-of-the-art overview regarding the development of 3D culture models, which allow for addressing research questions in models resembling the in vivo situation. In particular, we give an overview of the current developments of corneal 3D and organoid models, as well as 3D cell culture models of epithelia with goblet cells (conjunctiva and nasal cavity). The benefits and shortcomings of these cell culture models are discussed. As examples for pathogens related to ocular and nasal epithelia, we discuss infections caused by adenovirus and measles virus. In addition to pathogens, also external triggers such as allergens can cause rhinoconjunctivitis. These diseases exemplify the interconnections between the ocular surface and nasal epithelia in a molecular and clinical context. With a final translational section on optical coherence tomography (OCT), we provide an overview about the applicability of this technique in basic research and clinical ophthalmology. The techniques presented herein will be instrumental in further elucidating the functional interrelations and crosstalk between ocular and nasal epithelia.

scholarly journals Optimizations of In Vitro Mucus and Cell Culture Models to Better Predict In Vivo Gene Transfer in Pathological Lung Respiratory Airways: Cystic Fibrosis as an Example

Pharmaceutics ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 47
Author(s):  
Rosy Ghanem ◽  
Véronique Laurent ◽  
Philippe Roquefort ◽  
Tanguy Haute ◽  
Sophie Ramel ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Gene Therapy ◽  
Cell Culture ◽  
Gene Transfer ◽  
Gene Delivery ◽  
Culture Models ◽  
Transfer Strategies ◽  
Cell Culture Models

The respiratory epithelium can be affected by many diseases that could be treated using aerosol gene therapy. Among these, cystic fibrosis (CF) is a lethal inherited disease characterized by airways complications, which determine the life expectancy and the effectiveness of aerosolized treatments. Beside evaluations performed under in vivo settings, cell culture models mimicking in vivo pathophysiological conditions can provide complementary insights into the potential of gene transfer strategies. Such models must consider multiple parameters, following the rationale that proper gene transfer evaluations depend on whether they are performed under experimental conditions close to pathophysiological settings. In addition, the mucus layer, which covers the epithelial cells, constitutes a physical barrier for gene delivery, especially in diseases such as CF. Artificial mucus models featuring physical and biological properties similar to CF mucus allow determining the ability of gene transfer systems to effectively reach the underlying epithelium. In this review, we describe mucus and cellular models relevant for CF aerosol gene therapy, with a particular emphasis on mucus rheology. We strongly believe that combining multiple pathophysiological features in single complex cell culture models could help bridge the gaps between in vitro and in vivo settings, as well as viral and non-viral gene delivery strategies.

scholarly journals Selenomethionine-Dominated Selenium-Enriched Peanut Protein Ameliorates Alcohol-Induced Liver Disease in Mice by Suppressing Oxidative Stress

Foods ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2979
Author(s):  
Lin Gao ◽  
Jiawei Yuan ◽  
Yuhuan Cheng ◽  
Mengling Chen ◽  
Genhua Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Liver Disease ◽  
Therapeutic Agents ◽  
Peanut Protein ◽  
Culture Models ◽  
Therapeutic Properties ◽  
Cell Culture Models ◽  
Dependent Mechanism

Numerous natural compounds are considered as potential therapeutic agents against alcohol-induced liver disease (ALD). Research shows that selenium (Se) has a variety of bioactivities, including liver protecting ability. The present study based on in vitro cell culture models and in vivo mouse models was aimed at examining the contribution of selenomethionine (SeMet)-dominated Se-enriched peanut protein (SePP) to liver protection. SeMet and especially SePP reversed cell viability and cell death, inhibited ethanol induced CYP2E1 activation, decreased reactive oxygen species level, and restored GSH level. Hence, SeMet-dominated SePP alleviates alcohol-induced AML-12 cytotoxicity by suppressing oxidative stress. The p38-dependent mechanism was found to be responsible for SePP-induced Nrf-2 activation. Furthermore, supplementation with SePP and SeMet regulated lipid metabolism and reduced oxidative stress, minimizing liver damage in mice. Selenomethionine-dominated SePP possesses potential therapeutic properties and can be used to treat ALD through the suppression of oxidative stress.

Three-Dimensional Cell Culture Models of Hepatocellular Carcinoma — a Review

2021 ◽  
Author(s):  
Irmak Ayvaz ◽  
Dilara Sunay ◽  
Ece Sariyar ◽  
Esra Erdal ◽  
Zeynep Firtina Karagonlar
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Culture ◽  
Three Dimensional ◽  
Culture Models ◽  
Cell Culture Models ◽  
Dimensional Cell

scholarly journals 3D Cell Culture Models in COVID-19 Times: A Review of 3D Technologies to Understand and Accelerate Therapeutic Drug Discovery

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 602
Author(s):  
Guadalupe Tonantzin de Dios-Figueroa ◽  
Janette del Rocío Aguilera-Marquez ◽  
Tanya A. Camacho-Villegas ◽  
Pavel H. Lugo-Fabres
Keyword(s):  
Cell Culture ◽  
Drug Evaluation ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Therapeutic Drug ◽  
Emerging Viruses ◽  
Culture Models ◽  
Tissue Characteristics ◽  
Cell Culture Models

In the last decades, emerging viruses have become a worldwide concern. The fast and extensive spread of the disease caused by SARS-CoV-2 (COVID-19) has impacted the economy and human activity worldwide, highlighting the human vulnerability to infectious diseases and the need to develop and optimize technologies to tackle them. The three-dimensional (3D) cell culture models emulate major tissue characteristics such as the in vivo virus–host interactions. These systems may help to generate a quick response to confront new viruses, establish a reliable evaluation of the pathophysiology, and contribute to therapeutic drug evaluation in pandemic situations such as the one that humanity is living through today. This review describes different types of 3D cell culture models, such as spheroids, scaffolds, organoids, and organs-on-a-chip, that are used in virus research, including those used to understand the new severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2).

scholarly journals Initial Interaction of Rotavirus Strains with N-Acetylneuraminic (Sialic) Acid Residues on the Cell Surface Correlates with VP4 Genotype, Not Species of Origin

2002 ◽  
Vol 76 (8) ◽  
pp. 4087-4095 ◽  
Author(s):  
Max Ciarlet ◽  
Juan E. Ludert ◽  
Miren Iturriza-Gómara ◽  
Ferdinando Liprandi ◽  
James J. Gray ◽  
...  
Keyword(s):  
Cell Culture ◽  
Sialic Acid ◽  
Cell Surface ◽  
Correlation Coefficient ◽  
Molecular Basis ◽  
Human Origin ◽  
Culture Models ◽  
Initial Interaction ◽  
Cell Culture Models

ABSTRACT We examined 41 human and animal rotavirus strains representative of all known P genotypes for their dependency on cellular N-acetylneuraminic (sialic) acid (SA) residues for infectivity. Our results showed that all rotaviruses studied, whether of animal or human origin, belonging to P genotypes [1], [2], [3], and [7] depended on SA residues on the cell surface for efficient infectivity but that all human and animal rotavirus strains representative of the remaining known P genotypes were SA independent. The SA residue requirement for efficient infectivity did not change for reassortant rotavirus strains with altered VP4-VP7 combinations. The initial interaction of rotavirus strains with SA residues on the cell surface correlated with VP4 genotype specifity, not with species of origin or VP7 G serotype specificity (P = 0.001; r 2 = 1.00, Pearson's correlation coefficient). In addition to being a requirement for infectivity, the presence of SA residues on the cell surface is a requirement for efficient growth in cell culture; recognition of the association of specific P genotypes with the binding of rotavirus to SA residues will facilitate our understanding of the molecular basis of the early events of rotavirus-cell interactions in cell culture models and of pathogenicity in vivo.

scholarly journals Secretion of full-length Tau or Tau fragments in cell culture models. Propagation of Tau in vivo and in vitro

2018 ◽  
Vol 9 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Mar Pérez ◽  
Miguel Medina ◽  
Félix Hernández ◽  
Jesús Avila
Keyword(s):  
Cell Culture ◽  
Brain Regions ◽  
Tau Pathology ◽  
Protein Tau ◽  
Culture Models ◽  
Intracellular Aggregates ◽  
Cell Culture Models ◽  
The Brain

AbstractThe microtubule-associated protein Tau plays a crucial role in stabilizing neuronal microtubules. In Tauopathies, Tau loses its ability to bind microtubules, detach from them and forms intracellular aggregates. Increasing evidence in recent years supports the notion that Tau pathology spreading throughout the brain in AD and other Tauopathies is the consequence of the propagation of specific Tau species along neuroanatomically connected brain regions in a so-called “prion-like” manner. A number of steps are assumed to be involved in this process, including secretion, cellular uptake, transcellular transfer and/or seeding, although the precise mechanisms underlying propagation of Tau pathology are not fully understood yet. This review summarizes recent evidence on the nature of the specific Tau species that are propagated and the different mechanisms of Tau pathology spreading.

scholarly journals 3D Tissue Models as an Effective Tool for Studying Viruses and Vaccine Development

2021 ◽  
Vol 8 ◽  
Author(s):  
Nathan Lawko ◽  
Charlie Plaskasovitis ◽  
Carling Stokes ◽  
Laila Abelseth ◽  
Ian Fraser ◽  
...  
Keyword(s):  
Cell Culture ◽  
Drug Discovery ◽  
Vaccine Development ◽  
Rapid Development ◽  
3D Cell Culture ◽  
Host Cells ◽  
Culture Models ◽  
Cell Culture Models ◽  
Cell Cell

The recent SARS-CoV-2 outbreak has researchers working tirelessly to understand the virus' pathogenesis and develop an effective vaccine. The urgent need for rapid development and deployment of such a vaccine has illustrated the limitations of current practices, and it has highlighted the need for alternative models for early screening of such technologies. Traditional 2D cell culture does not accurately capture the effects of a physiologically relevant environment as they fail to promote appropriate cell-cell and cell-environment interactions. This inability to capture the intricacies of the in vivo microenvironment prevents 2D cell cultures from demonstrating the necessary properties of native tissues required for the standard infection mechanisms of the virus, thus contributing the high failure rate of drug discovery and vaccine development. 3D cell culture models can bridge the gap between conventional cell culture and in vivo models. Methods such as 3D bioprinting, spheroids, organoids, organ-on-chip platform, and rotating wall vessel bioreactors offer ways to produce physiologically relevant models by mimicking in vivo microarchitecture, chemical gradients, cell–cell interactions and cell–environment interactions. The field of viral biology currently uses 3D cell culture models to understand the interactions between viruses and host cells, which is crucial knowledge for vaccine development. In this review, we discuss how 3D cell culture models have been used to investigate disease pathologies for coronaviruses and other viruses such as Zika Virus, Hepatitis, and Influenza, and how they may apply to drug discovery and vaccine development.

scholarly journals An optimized, robust and reproducible protocol to generate well-differentiated primary nasal epithelial models from extremely premature infants

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Anke Martens ◽  
Gabriele Amann ◽  
Katy Schmidt ◽  
René Gaupmann ◽  
Bianca Böhm ◽  
...  
Keyword(s):  
Cell Culture ◽  
Premature Infants ◽  
Inflammatory Responses ◽  
Respiratory Infections ◽  
Successful Isolation ◽  
Culture Models ◽  
Well Differentiated ◽  
Air Liquid Interface ◽  
Cell Culture Models

AbstractExtremely premature infants are prone to severe respiratory infections, and the mechanisms underlying this exceptional susceptibility are largely unknown. Nasal epithelial cells (NEC) represent the first-line of defense and adult-derived ALI cell culture models show promising results in mimicking in vivo physiology. Therefore, the aim of this study was to develop a robust and reliable protocol for generating well-differentiated cell culture models from NECs of extremely premature infants. Nasal brushing was performed in 13 extremely premature infants at term corrected age and in 11 healthy adult controls to obtain NECs for differentiation at air-liquid interface (ALI). Differentiation was verified using imaging and functional analysis. Successful isolation and differentiation was achieved for 5 (38.5%) preterm and 5 (45.5%) adult samples. Preterm and adult ALI-cultures both showed well-differentiated morphology and ciliary function, however, preterm cultures required significantly longer cultivation times for acquiring full differentiation (44 ± 3.92 vs. 23 ± 1.83 days; p < 0.0001). Moreover, we observed that recent respiratory support may impair successful NECs isolation. Herewithin, we describe a safe, reliable and reproducible method to generate well-differentiated ALI-models from NECs of extremely premature infants. These models provide a valuable foundation for further studies regarding immunological and inflammatory responses and respiratory disorders in extremely premature infants.

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