High-content analysis of particulate matters-induced oxidative stress and organelle dysfunction in vitro

2019 ◽  
Vol 59 ◽  
pp. 263-274 ◽  
Author(s):  
Guanglei Wang ◽  
Xiaomei Zheng ◽  
Huawei Duan ◽  
Yufei Dai ◽  
Yong Niu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Content Analysis ◽  
Particulate Matters ◽  
High Content Analysis

scholarly journals Machine learning-assisted high-content analysis of pluripotent stem cell-derived embryos in vitro

2021 ◽  
Author(s):  
Jianying Guo ◽  
Peizhe Wang ◽  
Berna Sozen ◽  
Hui Qiu ◽  
Yonglin Zhu ◽  
...  
Keyword(s):  
Machine Learning ◽  
Content Analysis ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
High Content Analysis

High Content Analysis of an In Vitro Model for Metabolic Toxicity

2014 ◽  
Vol 19 (10) ◽  
pp. 1402-1408 ◽  
Author(s):  
Stephanie D. Cole ◽  
Janna S. Madren-Whalley ◽  
Albert P. Li ◽  
Russell Dorsey ◽  
Harry Salem
Keyword(s):  
Content Analysis ◽  
High Throughput ◽  
Metabolic Activation ◽  
Hepatic Metabolism ◽  
Cell Types ◽  
Multiple Organ ◽  
Compound Libraries ◽  
High Content Analysis ◽  
Calcein Am

In vitro models that accurately and rapidly assess hepatotoxicity and the effects of hepatic metabolism on nonliver cell types are needed by the U.S. Department of Defense and the pharmaceutical industry to screen compound libraries. Here, we report the first use of high content analysis on the Integrated Discrete Multiple Organ Co-Culture (IdMOC) system, a high-throughput method for such studies. We cultured 3T3-L1 cells in the presence and absence of primary human hepatocytes, and exposed the cultures to 4-aminophenol and cyclophosphamide, model toxicants that are respectively detoxified and activated by the liver. Following staining with calcein-AM, ethidium homodimer-1, and Hoechst 33342, high content analysis of the cultures revealed four cytotoxic endpoints: fluorescence intensities of calcein-AM and ethidium homodimer-1, nuclear area, and cell density. Using these endpoints, we observed that the cytotoxicity of 4-aminophenol in 3T3-L1 cells in co-culture was less than that observed for 3T3-L1 monocultures, consistent with the known detoxification of 4-aminophenol by hepatocytes. Conversely, cyclophosphamide cytotoxicity for 3T3-L1 cells was enhanced by co-culturing with hepatocytes, consistent with the known metabolic activation of this toxicant. The use of IdMOC plates combined with high content analysis is therefore a multi-endpoint, high-throughput capability for measuring the effects of metabolism on toxicity.

scholarly journals After In Vitro Digestion, Jackfruit Flake Affords Protection against Acrylamide-Induced Oxidative Damage

Molecules ◽  
2019 ◽  
Vol 24 (18) ◽  
pp. 3322 ◽  
Author(s):  
Daofeng Qu ◽  
Chu Liu ◽  
Mengxue Jiang ◽  
Lifang Feng ◽  
Yuewen Chen ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oxidative Damage ◽  
High Performance ◽  
In Vitro Digestion ◽  
Protective Effects ◽  
Physical Damage ◽  
Before And After ◽  
High Content Analysis ◽  
High Level

Some studies have demonstrated that acrylamide (AA) was correlated with oxidative stress, resulting in physical damage. The jackfruit flake was an immature pulp that contained a high level of antioxidant activity. This study aimed to assess the defensive efficacy of jackfruit flake in AA-induced oxidative stress before and after simulated gastrointestinal digestion. Our results indicate that the total polyphenol content of Jackfruit flake digest (Digestive products of jackfruit flake after gastrointestinal, JFG) was diminished; however, JFG had raised the relative antioxidant capacity compared to Jackfruit flake extract (JFE). Additionally, the results of High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) implied that a proportion of compounds were degraded/converted into other unknown and/or undetected metabolites. Further, by high content analysis (HCA) techniques, JFG markedly reduced cytotoxicity and excessive production of reactive oxygen species (ROS) in cells, thereby alleviating mitochondrial disorders. In this study, it may be converted active compounds after digestion that had preferable protective effects against AA-induced oxidative damage.

scholarly journals A Mechanistic High-Content Analysis Assay Using a Chimeric Androgen Receptor That Rapidly Characterizes Androgenic Chemicals

2020 ◽  
Vol 25 (7) ◽  
pp. 695-708
Author(s):  
Adam T. Szafran ◽  
Michael J. Bolt ◽  
Caroline E. Obkirchner ◽  
Maureen G. Mancini ◽  
Christine Helsen ◽  
...  
Keyword(s):  
Content Analysis ◽  
Androgen Receptor ◽  
Environmental Samples ◽  
Nuclear Translocation ◽  
Cell Model ◽  
Functional Assay ◽  
Chromatin Binding ◽  
Wide Range ◽  
High Content Analysis

Human health is at risk from environmental exposures to a wide range of chemical toxicants and endocrine-disrupting chemicals (EDCs). As part of understanding this risk, the U.S. Environmental Protection Agency (EPA) has been pursuing new high-throughput in vitro assays and computational models to characterize EDCs. EPA models have incorporated our high-content analysis–based green fluorescent protein estrogen receptor (GFP-ER): PRL-HeLa assay, which allows direct visualization of ER binding to DNA regulatory elements. Here, we characterize a modified functional assay based on the stable expression of a chimeric androgen receptor (ARER), wherein a region containing the native AR DNA-binding domain (DBD) was replaced with the ERα DBD (amino acids 183–254). We demonstrate that the AR agonist dihydrotestosterone induces GFP-ARER nuclear translocation, PRL promoter binding, and transcriptional activity at physiologically relevant concentrations (<1 nM). In contrast, the AR antagonist bicalutamide induces only nuclear translocation of the GFP-ARER receptor (at μM concentrations). Estradiol also fails to induce visible chromatin binding, indicating androgen specificity. In a screen of reference chemicals from the EPA and the Agency for Toxic Substances and Disease Registry, the GFP-ARER cell model identified and mechanistically grouped activity by known (anti-)androgens based on the ability to induce nuclear translocation and/or chromatin binding. Finally, the cell model was used to identify potential (anti-)androgens in environmental samples in collaboration with the Houston Ship Channel/Galveston Bay Texas A&M University EPA Superfund Research Program. Based on these data, the chromatin-binding, in vitro assay–based GFP-ARER model represents a selective tool for rapidly identifying androgenic activity associated with drugs, chemicals, and environmental samples.

Simultaneous in situ visualization and quantitation of dual antigens adsorbed on adjuvants using high content analysis

Nanomedicine ◽  
2019 ◽  
Vol 14 (19) ◽  
pp. 2535-2548 ◽  
Author(s):  
Zhigang Zhang ◽  
Tianying Zhang ◽  
Lu Cao ◽  
Xin Wang ◽  
Jiali Cao ◽  
...  
Keyword(s):  
Content Analysis ◽  
Human Papillomavirus Vaccine ◽  
In Situ Method ◽  
Vaccine Antigens ◽  
High Content Analysis ◽  
Papillomavirus Vaccine ◽  
Simultaneous Visualization ◽  
Good Agreement

Aim: Traditional antigenicity assay requires antigen recovery from the particulate adjuvants prior to analysis. An in situ method was developed for interrogating vaccine antigens with monoclonal antibodies while being adsorbed on adjuvants. Materials & methods: The fluorescence imaging-based high content analysis was used to visualize the antigen distribution on adjuvant agglomerates and to analyze the antigenicity for adsorbed antigens. Results: Simultaneous visualization and quantitation were achieved for dual antigens in a bivalent human papillomavirus vaccine with uniquely labeled antibodies. Good agreement was observed between the in situ multiplexed assays with well-established sandwich enzyme-linked immunosorbent assays. Conclusion: The streamlined procedures and the amenability for multiplexing make the in situ antigenicity analysis a favorable choice for in vitro functional assessment of bionanoparticles as vaccine antigens.

scholarly journals 3D Cell-Based Assays for Drug Screens: Challenges in Imaging, Image Analysis, and High-Content Analysis

2019 ◽  
Vol 24 (6) ◽  
pp. 615-627 ◽  
Author(s):  
Tijmen H. Booij ◽  
Leo S. Price ◽  
Erik H. J. Danen
Keyword(s):  
Content Analysis ◽  
Drug Discovery ◽  
High Throughput Screening ◽  
3D Cell Culture ◽  
Automated Analysis ◽  
In Vivo Models ◽  
Current Switch ◽  
High Content Analysis

The introduction of more relevant cell models in early preclinical drug discovery, combined with high-content imaging and automated analysis, is expected to increase the quality of compounds progressing to preclinical stages in the drug development pipeline. In this review we discuss the current switch to more relevant 3D cell culture models and associated challenges for high-throughput screening and high-content analysis. We propose that overcoming these challenges will enable front-loading the drug discovery pipeline with better biology, extracting the most from that biology, and, in general, improving translation between in vitro and in vivo models. This is expected to reduce the proportion of compounds that fail in vivo testing due to a lack of efficacy or to toxicity.

scholarly journals High Content Analysis of Human Pluripotent Stem Cell Derived Hepatocytes Reveals Drug Induced Steatosis and Phospholipidosis

2016 ◽  
Vol 2016 ◽  
pp. 1-14 ◽  
Author(s):  
Arvind Pradip ◽  
Daniella Steel ◽  
Susanna Jacobsson ◽  
Gustav Holmgren ◽  
Magnus Ingelman-Sundberg ◽  
...  
Keyword(s):  
Content Analysis ◽  
Toxicity Testing ◽  
Published Data ◽  
Drug Induced ◽  
Attrition Rates ◽  
Pure Cultures ◽  
High Content Analysis ◽  
Approved Drugs

Hepatotoxicity is one of the most cited reasons for withdrawal of approved drugs from the market. The use of nonclinically relevantin vitroandin vivotesting systems contributes to the high attrition rates. Recent advances in differentiating human induced pluripotent stem cells (hiPSCs) into pure cultures of hepatocyte-like cells expressing functional drug metabolizing enzymes open up possibilities for novel, more relevant human cell based toxicity models. The present study aimed to investigate the use of hiPSC derived hepatocytes for conducting mechanistic toxicity testing by image based high content analysis (HCA). The hiPSC derived hepatocytes were exposed to drugs known to cause hepatotoxicity through steatosis and phospholipidosis, measuring several endpoints representing different mechanisms involved in drug induced hepatotoxicity. The hiPSC derived hepatocytes were benchmarked to the HepG2 cell line and generated robust HCA data with low imprecision between plates and batches. The different parameters measured were detected at subcytotoxic concentrations and the order of which the compounds were categorized (as severe, moderate, mild, or nontoxic) based on the degree of injury at isomolar concentration corresponded to previously published data. Taken together, the present study shows how hiPSC derived hepatocytes can be used as a platform for screening drug induced hepatotoxicity by HCA.

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