scholarly journals The Effect of Primary Cancer Cell Culture Models on the Results of Drug Chemosensitivity Assays: The Application of Perfusion Microbioreactor System as Cell Culture Vessel

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Chia-Hsun Hsieh ◽  
Yi-Dao Chen ◽  
Shiang-Fu Huang ◽  
Hung-Ming Wang ◽  
Min-Hsien Wu
Keyword(s):  
Cell Culture ◽  
Metabolic Activity ◽  
Culture Condition ◽  
3D Culture ◽  
Culture Vessel ◽  
Chemosensitivity Assay ◽  
Culture Models ◽  
Tumor Level ◽  
Chemosensitivity Assays ◽  
The Impact

To precisely and faithfully perform cell-based drug chemosensitivity assays, a well-defined and biologically relevant culture condition is required. For the former, a perfusion microbioreactor system capable of providing a stable culture condition was adopted. For the latter, however, little is known about the impact of culture models on the physiology and chemosensitivity assay results of primary oral cavity cancer cells. To address the issues, experiments were performed. Results showed that minor environmental pH change could significantly affect the metabolic activity of cells, demonstrating the importance of stable culture condition for such assays. Moreover, the culture models could also significantly influence the metabolic activity and proliferation of cells. Furthermore, the choice of culture models might lead to different outcomes of chemosensitivity assays. Compared with the similar test based on tumor-level assays, the spheroid model could overestimate the drug resistance of cells to cisplatin, whereas the 2D and 3D culture models might overestimate the chemosensitivity of cells to such anticancer drug. In this study, the 3D culture models with same cell density as that in tumor samples showed comparable chemosensitivity assay results as the tumor-level assays. Overall, this study has provided some fundamental information for establishing a precise and faithful drug chemosensitivity assay.

scholarly journals Factors to consider when interrogating 3D culture models with plate readers or automated microscopes

2021 ◽  
Author(s):  
Terry Riss ◽  
O. Joseph Trask
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Culture ◽  
Fluorescent Imaging ◽  
Culture Models ◽  
Assay Methods ◽  
Diffusion Rates ◽  
Cell Culture Models ◽  
Dimensional Cell ◽  
Cells Cultured

AbstractAlong with the increased use of more physiologically relevant three-dimensional cell culture models comes the responsibility of researchers to validate new assay methods that measure events in structures that are physically larger and more complex compared to monolayers of cells. It should not be assumed that assays designed using monolayers of cells will work for cells cultured as larger three-dimensional masses. The size and barriers for penetration of molecules through the layers of cells result in a different microenvironment for the cells in the outer layer compared to the center of three-dimensional structures. Diffusion rates for nutrients and oxygen may limit metabolic activity which is often measured as a marker for cell viability. For assays that lyse cells, the penetration of reagents to achieve uniform cell lysis must be considered. For live cell fluorescent imaging assays, the diffusion of fluorescent probes and penetration of photons of light for probe excitation and fluorescent emission must be considered. This review will provide an overview of factors to consider when implementing assays to interrogate three dimensional cell culture models.

scholarly journals Applying phasor approach analysis of multiphoton FLIM measurements to probe the metabolic activity of three-dimensional in vitro cell culture models

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Pirmin H. Lakner ◽  
Michael G. Monaghan ◽  
Yvonne Möller ◽  
Monilola A. Olayioye ◽  
Katja Schenke-Layland
Keyword(s):  
Cell Culture ◽  
Metabolic Activity ◽  
Three Dimensional ◽  
In Vitro Cell Culture ◽  
Culture Models ◽  
Cell Culture Models

scholarly journals Cellular Effects of T-2 Toxin on Primary Hepatic Cell Culture Models of Chickens

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 46 ◽  
Author(s):  
Máté Mackei ◽  
Kata Orbán ◽  
Andor Molnár ◽  
László Pál ◽  
Károly Dublecz ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Culture ◽  
Cell Cultures ◽  
Metabolic Activity ◽  
Incubation Time ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Cellular Effects ◽  
Culture Models ◽  
Cell Culture Models

Trichothecene mycotoxins such as T-2 toxin cause severe problems for agriculture, as well as for veterinary medicine. As liver is one of the key organs in metabolism, the main aim of our study was to investigate the immunomodulatory and cytotoxic effects of T-2 toxin, using primary hepatocyte mono-culture and hepatocyte—nonparenchymal cell (predominantly Kupffer cell) co-culture models of chicken. Cultures were exposed to 10 (T10 group), 100 (T100 group) and 1000 (T1000 group) nmol/L T-2 toxin treatment for 8 or 24 h. Alterations of cellular metabolic activity, the production of reactive oxygen species (extracellular H2O2), heat shock protein 70 (HSP70), and the concentration of different inflammatory cytokines such as interleukin (IL-)6 and IL-8 were investigated. Metabolic activity was intensely decreased by T-2 toxin administration in all of the cell culture models, in every applied concentration and incubation time. Concentrations of HSP70 and IL-8 were significantly increased in hepatocyte mono-cultures exposed to higher T-2 toxin levels (both in T100 and T1000 groups for HSP70 and in T1000 group for IL-8, respectively) compared to controls after 24 h incubation. Similarly, IL-6 levels were also significantly elevated in the T100 and T1000 groups in both of mono- and co-cultures, but only after 8 h of incubation time. In spite of the general harmful effects of T-2 toxin treatment, no significant differences were observed on reactive oxygen species production. Furthermore, the two cell culture models showed different levels of H2O2, HSP70, and IL-8 concentrations independently of T-2 toxin supplementation. In conclusion, the established primary cell cultures derived from chicken proved to be proper models to study the specific molecular effects caused by T-2 toxin. Metabolic activity and immune status of the different examined cell cultures were intensively affected; however, no changes were found in H2O2 levels.

Effect of cell culture models on the evaluation of anticancer activity and mechanism analysis of the potential bioactive compound, iturin A, produced by Bacillus subtilis

2019 ◽  
Vol 10 (3) ◽  
pp. 1478-1489 ◽  
Author(s):  
Haobin Zhao ◽  
Xixi Zhao ◽  
Shuzhen Lei ◽  
Yawen Zhang ◽  
Dongyan Shao ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bacillus Subtilis ◽  
3D Culture ◽  
Bioactive Compound ◽  
Mechanism Analysis ◽  
Iturin A ◽  
Culture Cells ◽  
Culture Models ◽  
Cell Culture Models ◽  
Cells Cultured

Compared to 2D culture, cells cultured in 3D culture were more resistant to iturin A from Bacillus.

scholarly journals Design, characterization, and modeling of microcirculation systems with integrated oxygenators

2016 ◽  
Vol 5 (1) ◽  
pp. 221-228 ◽  
Author(s):  
Mathias Busek ◽  
Stefan Gruenzner ◽  
Tobias Steege ◽  
Florian Schmieder ◽  
Udo Klotzbach ◽  
...  
Keyword(s):  
Cell Culture ◽  
Gas Permeability ◽  
Basic Research ◽  
Microfluidic System ◽  
Animal Testing ◽  
Permeable Membrane ◽  
Micro Particle Image Velocimetry ◽  
Hypoxic Conditions ◽  
Culture Models ◽  
The Impact

Abstract. Here, we describe a microfluidic system for hypoxia assays on human cell culture models. These systems are developed to replace or reduce animal testing in biomedical basic research. The presented system uses a gas-permeable membrane as a gas–liquid interface and a micropump for media actuation to influence the oxygen content in two cell culture chambers. To apply well-defined hypoxic conditions to the cells, a good understanding of the mass transport phenomena is necessary. Therefore, a complete network model of the microfluidic system is presented. This model is validated by means of micro-particle image velocimetry (µPIV) and optical oxygen measurement with fluorescence lifetime detection. Finally, the impact of several process parameters, e.g., the gas permeability of the pump, is discussed using the developed model.

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 NSAID therapy effects on healing of bone, tendon, and the enthesis

2013 ◽  
Vol 115 (6) ◽  
pp. 892-899 ◽  
Author(s):  
Bailey Su ◽  
J. Patrick O'Connor
Keyword(s):  
Cell Culture ◽  
Nsaid Therapy ◽  
Pharmacological Properties ◽  
Physiological Processes ◽  
Cyclooxygenase Activity ◽  
Culture Models ◽  
Dosing Regimens ◽  
Inflammatory Drugs ◽  
The Impact ◽  
Cell Culture Models

Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used for the treatment of skeletal injuries. The ability of NSAIDs to reduce pain and inflammation is well-established. However, the effects of NSAID therapy on healing of skeletal injuries is less defined. NSAIDs inhibit cyclooxygenase activity to reduce synthesis of prostaglandins, which are proinflammatory, lipid-signaling molecules. Inhibition of cyclooxygenase activity can impact many physiological processes. The effects of NSAID therapy on healing of bone, tendon, and the tendon-to-bone junction (enthesis) have been studied in animal and cell culture models, but human studies are few. Use of different NSAIDs with different pharmacological properties, differences in dosing regimens, and differences in study models and outcome measures have complicated comparisons between studies. In this review, we summarize the mechanisms by which bone, tendon, and enthesis healing occurs, and describe the effects of NSAID therapy on each of these processes. Determining the impact of NSAID therapy on healing of skeletal tissues will enable clinicians to appropriately manage the patient's condition and improve healing outcomes.

scholarly journals 3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs

Cancers ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 190
Author(s):  
Mélanie A. G. Barbosa ◽  
Cristina P. R. Xavier ◽  
Rúben F. Pereira ◽  
Vilma Petrikaitė ◽  
M. Helena Vasconcelos
Keyword(s):  
Cell Culture ◽  
Tumor Microenvironment ◽  
Tumor Cells ◽  
3D Cell Culture ◽  
Drug Efficacy ◽  
3D Models ◽  
Laboratory Animals ◽  
Culture Models ◽  
The Impact ◽  
Cell Culture Models

Today, innovative three-dimensional (3D) cell culture models have been proposed as viable and biomimetic alternatives for initial drug screening, allowing the improvement of the efficiency of drug development. These models are gaining popularity, given their ability to reproduce key aspects of the tumor microenvironment, concerning the 3D tumor architecture as well as the interactions of tumor cells with the extracellular matrix and surrounding non-tumor cells. The development of accurate 3D models may become beneficial to decrease the use of laboratory animals in scientific research, in accordance with the European Union’s regulation on the 3R rule (Replacement, Reduction, Refinement). This review focuses on the impact of 3D cell culture models on cancer research, discussing their advantages, limitations, and compatibility with high-throughput screenings and automated systems. An insight is also given on the adequacy of the available readouts for the interpretation of the data obtained from the 3D cell culture models. Importantly, we also emphasize the need for the incorporation of additional and complementary microenvironment elements on the design of 3D cell culture models, towards improved predictive value of drug efficacy.

scholarly journals Integrative data analysis predicts YY1 as a Cis-regulator in the 3D Cell Culture Models of MCF10A at the Stiffness Level of High Mammographic Density

2018 ◽  
Author(s):  
Qingsu Cheng ◽  
Mina Khoshdeli ◽  
Chongzhi Zang ◽  
Bahram Parvin
Keyword(s):  
Cell Culture ◽  
Transcription Factors ◽  
Mammographic Density ◽  
Regulation Of Gene Expression ◽  
3D Culture ◽  
3D Cell Culture ◽  
Transcriptome Data ◽  
High Mammographic Density ◽  
Culture Models ◽  
Cell Culture Models

AbstractPrevious studies have shown that in 3D cell culture models of human mammary cells (HMEC) (i) colony organizations are heterogeneous, and (ii) ERBB2 is overexpressed in MCF10A when the stiffness of the microenvironment is increased to that of high mammographic density (MD). The goal of the current study is to identify transcription factors that regulate processes associated with the increased stiffness of the microenvironment. Two HMEC premalignant lines of MCF7 and 184A1 are cultured in 3D, colonies are imaged using confocal microscopy, and colony organizations and heterogeneity are quantified as a function of the stiffness of the microenvironment. In parallel and surrogate assays, colony organizations are profiled by transcriptomics. Transcriptome data are enriched by correlative analysis with the computed morphometric indices, from 3D culture, and a subset of transcriptome data is selected. This subset is then processed with Model-based Analysis of Regulation of Gene Expression (MARGE) and publicly available ChIP-seq data to predict regulatory transcription factors. The integrative analysis indicated that YY1 regulates ERBB2 in the 3D cell culture of MCF10A when the stiffness of the microenvironment is increased to that of high MD. Subsequent experimental validation confirmed that YY1 is only expressed at the high stiffness value of the microenvironment concomitant with the overexpression of ERBB2 in MCF10A. Furthermore, using ERBB2 positive SKBR3 cell line, co-expression of YY1 and ERBB2 is absent, which indicates that YY1 regulates tumorigenicity through multiple pathways.Author’s summaryMCF10A is a premalignant immortalized human mammary cell that has been isolated from a patient with fibrocystic and lost several barriers toward transformation. In an earlier study, we showed that ERBB2 is upregulated in 3D cultures of MCF10A when the stiffness of the microenvironment is increased to that of high mammographic density. Here, we leverage publicly available ChIP-seq data to predict and validate the cis-regulator of ERBB2. Our integrated experimental and computation protocol provides a pathway for elucidating regulators that can potentially be targeted for intervention.

scholarly journals Brilacidin Demonstrates Inhibition of SARS-CoV-2 in Cell Culture

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 271 ◽  
Author(s):  
Allison Bakovic ◽  
Kenneth Risner ◽  
Nishank Bhalla ◽  
Farhang Alem ◽  
Theresa L. Chang ◽  
...  
Keyword(s):  
Cell Culture ◽  
Antiviral Activity ◽  
Cell Line ◽  
Viral Entry ◽  
De Novo ◽  
Biological Properties ◽  
Defense Proteins ◽  
Culture Models ◽  
Different Strains ◽  
The Impact

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the newly emergent causative agent of coronavirus disease-19 (COVID-19), has resulted in more than two million deaths worldwide since it was first detected in 2019. There is a critical global need for therapeutic intervention strategies that can be deployed to safely treat COVID-19 disease and reduce associated morbidity and mortality. Increasing evidence shows that both natural and synthetic antimicrobial peptides (AMPs), also referred to as Host Defense Proteins/Peptides (HDPs), can inhibit SARS-CoV-2, paving the way for the potential clinical use of these molecules as therapeutic options. In this manuscript, we describe the potent antiviral activity exerted by brilacidin—a de novo designed synthetic small molecule that captures the biological properties of HDPs—on SARS-CoV-2 in a human lung cell line (Calu-3) and a monkey cell line (Vero). These data suggest that SARS-CoV-2 inhibition in these cell culture models is likely to be a result of the impact of brilacidin on viral entry and its disruption of viral integrity. Brilacidin demonstrated synergistic antiviral activity when combined with remdesivir. Collectively, our data demonstrate that brilacidin exerts potent inhibition of SARS-CoV-2 against different strains of the virus in cell culture.

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