Download or read book In Vitro 3D Fluorescent Cell based Assay Reporting Gene Regulation for High throughput Drug Screening written by You Li and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Traditional cell-based assays have a high failure rate of identifying effective compounds in drug discovery. To address this problem, recent drug research has focused on engineering cancer cells that can indicate specific drug molecular mechanisms of action and developing cancer models that can more accurately represent tumor physiology. Survivin and telomerase reverse transcriptase (TERT), two important cancer biomarkers overexpressed in 85% of tumor tissues, play crucial roles in anti-apoptosis and unlimited cell proliferation in carcinogenesis, as well as drug resistance in cancer therapy. In this research, we have developed survivin and TERT reporter gene assays for high-throughput screening (HTS) of drugs downregulating these genes as anticancer therapeutics. Cancer cells were engineered to express fluorescent proteins controlled by TERT and survivin promoters, respectively, and were cultured in 3-dimensional (3D) scaffolds in microbioreactors to investigate drug effects by monitoring changes in culture fluorescence in real time. The novel 3-D assays offered molecular and physiological predictions of gene-specific responses to drugs, thus facilitating cancer drug discovery and development. Using survivin as the target, a dual-color fluorescent reporter gene assay performed in 3-D culture was developed. To demonstrate the drug's specific effect on the survivin promoter, MCF-7 breast cancer cells were engineered to express enhanced green fluorescent protein (EGFP) driven by a survivin promoter. The cell also expressed red fluorescent protein (RFP) under the control of a cytomegalovirus promoter as an internal control and to indicate cell number. The specific survivin expression was assessed by the green fluorescence normalized to the red fluorescence, which was used as an indicator to assess drug’s regulation on survivin. The internal control compensated for the variations in cell number, cell culture environment, fluorescent protein property, detection setting, and even for the reading interference from tested compounds, resulting in a more accurate evaluation of drug effect. The assay correctly detected the downregulation and upregulation of several anticancer compounds and demonstrated its reliability for high-throughput screening of survivin-targeted drugs. In this work, a 3-D cancer cell-based hTERT promoter assay was also developed for anticancer drug screening. Different lengths of the hTERT promoter were cloned with EGFP as reporting cassettes and were introduced in multiple cancer cell lines. The selected MCF7 reporter cell was able to correctly respond to the known hTERT inhibitors, pristimerin, epigallocatechin gallate and n-butylidenephthalide. The reporter cells were cultured on 3-D scaffolds to resemble in vivo tumor physiology and present in vivo like responses. The 3-D hTERT promoter assay was verified by pristimerin, and subsequently, other five anticancer compounds, including doxorubicin, cisplatin, paclitaxel, blasticidin, and tamoxifen, were tested for their effects on hTERT. The developed assay achieved an overall accuracy rate of over 80%, which demonstrated the feasibility of using 3D culture and EGFP as a reporter to screen hTERT inhibitors. In addition to the development of cancer cell-based assays, hTERT as a pluripotency marker to predict embryotoxic compounds during tissue development was also evaluated. The hTERT-EGFP reporting cassettes were introduced into mouse embryonic stem cells D3. The reportability was assessed by the corelated expression changes of EGFP and endogenous TERT during neural differentiation and after receiving toxicant stimuli. Our results showed that EGFP was able to indicate drug effect on human TERT expression level during ESC differentiation, which, however, was not always consistent with internal mouse TERT (mTERT) due to species difference. The results confirmed that drug effects on TERT were species-specific, especially at the beginning of differentiation. In conclusion, hTERT in mESCs reacted to embryotoxic compounds and the engineered mESC-hTERT-EGFP cells could be used for human teratogen prediction.
Download or read book Cell Based Assays for High Throughput Screening written by Paul A. Clemons and published by Humana Press. This book was released on 2014-11-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: As the use of high-throughput screening expands and creates more interest in the academic community, the need for detailed reference materials becomes ever more pressing. Cell-Based Assays for High-Throughput Screening: Methods and Protocols aims to fill an important part of this need by providing an easily accessible reference volume for cell-based phenotypic screening. Leading researchers in the field contribute state-of-the-art methods with actionable protocols covering four major areas of study: model biological systems, screening modalities and assay systems, detection technologies, and approaches to data analysis. Written in the highly successful Methods in Molecular BiologyTM series format, each chapter includes a brief introduction to the subject, lists of necessary materials and reagents, step-by-step laboratory protocols, and a Notes section detailing tips on troubleshooting and avoiding known pitfalls. Cutting-edge and easy-to-use, Cell-Based Assays for High-Throughput Screening: Methods and Protocols presents an overview of relevant approaches, enabling the direct application of existing methods to new discoveries while also inspiring researchers to approach their screening projects in a conceptually modular fashion, enhancing the power to discover through new combinations of existing approaches.
Download or read book Chemical Genomics written by Haian Fu and published by Cambridge University Press. This book was released on 2012-02-13 with total page 359 pages. Available in PDF, EPUB and Kindle. Book excerpt: Advances in chemistry, biology and genomics coupled with laboratory automation and computational technologies have led to the rapid emergence of the multidisciplinary field of chemical genomics. This edited text, with contributions from experts in the field, discusses the new techniques and applications that help further the study of chemical genomics. The beginning chapters provide an overview of the basic principles of chemical biology and chemical genomics. This is followed by a technical section that describes the sources of small-molecule chemicals; the basics of high-throughput screening technologies; and various bioassays for biochemical-, cellular- and organism-based screens. The final chapters connect the chemical genomics field with personalized medicine and the druggable genome for future discovery of new therapeutics. This book will be valuable to researchers, professionals and graduate students in many fields, including biology, biomedicine and chemistry.
Download or read book Phenotypic Drug Discovery written by Beverley Isherwood and published by Royal Society of Chemistry. This book was released on 2020-12-09 with total page 273 pages. Available in PDF, EPUB and Kindle. Book excerpt: Phenotypic drug discovery has been highlighted in the past decade as an important strategy in the discovery of new medical entities. How many marketed drugs are derived from phenotypic screens? From the most recent examples, what were the factors enabling target identification and validation? This book answers these questions by elaborating on fundamental capabilities required for phenotypic drug discovery and using case studies to illustrate approaches and key success factors. Written and edited by experienced practitioners from both industry and academia, this publication will equip researchers with a thought-provoking guide to the application and future development of contemporary phenotypic drug discovery for clinical success.
Download or read book Drug Discovery and Development written by Omboon Vallisuta and published by BoD – Books on Demand. This book was released on 2015-06-03 with total page 332 pages. Available in PDF, EPUB and Kindle. Book excerpt: It is very important for scientists all over the globe to enhance drug discovery research for better human health. This book demonstrates that various expertise are essential for drug discovery including synthetic or natural drugs, clinical pharmacology, receptor identification, drug metabolism, pharmacodynamic and pharmacokinetic research. The following 5 sections cover diverse chapter topics in drug discovery: Natural Products as Sources of Leading Molecules in Drug Discovery; Oncology and Drug Discovery; Receptors Involvement in Drug Discovery; Management and Development of Drugs against Infectious Diseases; Advanced Methodology.
Download or read book Special Topics in Drug Discovery written by Taosheng Chen and published by BoD – Books on Demand. This book was released on 2016-11-30 with total page 192 pages. Available in PDF, EPUB and Kindle. Book excerpt: Drug discovery involves multiple disciplines, technologies, and approaches. This book selects important topics related to drug discovery, including emerging tool (Chapter 1), cutting-edge approaches (Chapters 2, 3, and 4), examples of specific therapeutic area (Chapter 5), quality control in drug development (Chapter 6), and job and career opportunities in the pharmaceutical sector, a topic rarely covered by other books (Chapter 7). This book draws knowledge from experts actively involved in different areas of drug discovery from both industrial and academic settings. We hope that this book will facilitate your efforts in drug discovery.
Download or read book Make Life Visible written by Yoshiaki Toyama and published by Springer Nature. This book was released on 2019-10-02 with total page 292 pages. Available in PDF, EPUB and Kindle. Book excerpt: This open access book describes marked advances in imaging technology that have enabled the visualization of phenomena in ways formerly believed to be completelyimpossible. These technologies have made major contributions to the elucidation of the pathology of diseases as well as to their diagnosis and therapy. The volume presents various studies from molecular imaging to clinical imaging. It also focuses on innovative, creative, advanced research that gives full play to imaging technology inthe broad sense, while exploring cross-disciplinary areas in which individual research fields interact and pursuing the development of new techniques where they fuse together. The book is separated into three parts, the first of which addresses the topic of visualizing and controlling molecules for life. Th e second part is devoted to imaging of disease mechanisms, while the final part comprises studies on the application of imaging technologies to diagnosis and therapy. Th e book contains the proceedings of the 12th Uehara International Symposium 2017, “Make Life Visible” sponsored by the Uehara Memorial Foundation and held from June 12 to 14, 2017. It is written by leading scientists in the field and is an open access publication under a CC BY 4.0 license.
Download or read book High Throughput Screening in Drug Discovery written by Jörg Hüser and published by John Wiley & Sons. This book was released on 2006-12-13 with total page 362 pages. Available in PDF, EPUB and Kindle. Book excerpt: Backed by leading authorities, this is a professional guide to successful compound screening in pharmaceutical research and chemical biology, including the chemoinformatic tools needed for correct data evaluation. Chapter authors from leading pharmaceutical companies as well as from Harvard University discuss such factors as chemical genetics, binding, cell-based and biochemical assays, the efficient use of compound libraries and data mining using cell-based assay results. For both academics and professionals in the pharma and biotech industries working on small molecule screening.
Download or read book Angiogenesis Assays written by Carolyn A. Staton and published by John Wiley & Sons. This book was released on 2007-01-11 with total page 410 pages. Available in PDF, EPUB and Kindle. Book excerpt: Angiogenesis, the development of new blood vessels from the existing vasculature, is essential for physiological growth and over 18,000 research articles have been published describing the role of angiogenesis in over 70 different diseases, including cancer, diabetic retinopathy, rheumatoid arthritis and psoriasis. One of the most important technical challenges in such studies has been finding suitable methods for assessing the effects of regulators of eh angiogenic response. While increasing numbers of angiogenesis assays are being described both in vitro and in vivo, it is often still necessary to use a combination of assays to identify the cellular and molecular events in angiogenesis and the full range of effects of a given test protein. Although the endothelial cell - its migration, proliferation, differentiation and structural rearrangement - is central to the angiogenic process, it is not the only cell type involved. the supporting cells, the extracellular matrix and the circulating blood with its cellular and humoral components also contribute. In this book, experts in the use of a diverse range of assays outline key components of these and give a critical appraisal of their strengths and weaknesses. Examples include assays for the proliferation, migration and differentiation of endothelial cells in vitro, vessel outgrowth from organ cultures, assessment of endothelial and mural cell interactions, and such in vivo assays as the chick chorioallantoic membrane, zebrafish, corneal, chamber and tumour angiogenesis models. These are followed by a critical analysis of the biological end-points currently being used in clinical trials to assess the clinical efficacy of anti-angiogenic drugs, which leads into a discussion of the direction future studies should take. This valuable book is of interest to research scientists currently working on angiogenesis in both the academic community and in the biotechnology and pharmaceutical industries. Relevant disciplines include cell and molecular biology, oncology, cardiovascular research, biotechnology, pharmacology, pathology and physiology.
Download or read book Retinoid Signaling Pathways written by and published by Academic Press. This book was released on 2020-04-29 with total page 612 pages. Available in PDF, EPUB and Kindle. Book excerpt: Retinoid Signaling Pathways, Volume 637, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Sections in this release include The chemistry and biochemistry of Vitamin A and its natural derivative, Biosynthesis of retinoic acids, Biodegration of retinoic acids mediated by retinoid binding proteins, Retinoic acid homeostasis, Cryo Electron Microscopy to study retinol update via the STRA6 receptor, Immuno-detection of retinoic acid synthesis enzymes in the brain, classical pathway of gene regulation by retinoids, Protein-protein interactions in the regulation of retinoid acid receptors activity, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series Includes the latest information on retinoid signaling pathways
Download or read book Cell based Biosensors written by Qingjun Liu and published by Artech House. This book was released on 2009-10 with total page 291 pages. Available in PDF, EPUB and Kindle. Book excerpt: Written by recognized experts the field, this leading-edge resource is the first book to systematically introduce the concept, technology, and development of cell-based biosensors. You find details on the latest cell-based biosensor models and novel micro-structure biosensor techniques. Taking an interdisciplinary approach, this unique volume presents the latest innovative applications of cell-based biosensors in a variety of biomedical fields. The book also explores future trends of cell-based biosensors, including integrated chips, nanotechnology and microfluidics. Over 140 illustrations help clarify key topics throughout the book.
Download or read book 3D Cell Based Biosensors in Drug Discovery Programs written by William S. Kisaalita and published by CRC Press. This book was released on 2010-06-22 with total page 404 pages. Available in PDF, EPUB and Kindle. Book excerpt: Advances in genomics and combinatorial chemistry during the past two decades inspired innovative technologies and changes in the discovery and pre-clinical development paradigm with the goal of accelerating the process of bringing therapeutic drugs to market. Written by William Kisaalita, one of the foremost experts in this field, 3D Cell-Based Bio
Download or read book Tumor Organoids written by Shay Soker and published by Humana Press. This book was released on 2017-10-20 with total page 225 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cancer cell biology research in general, and anti-cancer drug development specifically, still relies on standard cell culture techniques that place the cells in an unnatural environment. As a consequence, growing tumor cells in plastic dishes places a selective pressure that substantially alters their original molecular and phenotypic properties.The emerging field of regenerative medicine has developed bioengineered tissue platforms that can better mimic the structure and cellular heterogeneity of in vivo tissue, and are suitable for tumor bioengineering research. Microengineering technologies have resulted in advanced methods for creating and culturing 3-D human tissue. By encapsulating the respective cell type or combining several cell types to form tissues, these model organs can be viable for longer periods of time and are cultured to develop functional properties similar to native tissues. This approach recapitulates the dynamic role of cell–cell, cell–ECM, and mechanical interactions inside the tumor. Further incorporation of cells representative of the tumor stroma, such as endothelial cells (EC) and tumor fibroblasts, can mimic the in vivo tumor microenvironment. Collectively, bioengineered tumors create an important resource for the in vitro study of tumor growth in 3D including tumor biomechanics and the effects of anti-cancer drugs on 3D tumor tissue. These technologies have the potential to overcome current limitations to genetic and histological tumor classification and development of personalized therapies.
Download or read book Endothelial Cell Culture written by Roy Bicknell and published by Cambridge University Press. This book was released on 1996-09-28 with total page 156 pages. Available in PDF, EPUB and Kindle. Book excerpt: The aim of the Handbooks in Practical Animal Cell Biology is to provide practical workbooks for those involved in primary cell culture. Each volume addresses a different cell lineage, and contains an introductory section followed by individual chapters on the culture of specific differentiated cell types. The authors of each chapter are leading researchers in their fields and use their first-hand experience to present reliable techniques in a clear and thorough manner. Endothelial Cell Culture contains chapters on endothelial cells derived from 1) lung, 2) bone marrow, 3) brain, 4) mammary glands, 5) skin, 6) adipose tissue, 7) female reproductive system, and 8) synovium.
Download or read book Bioelectrochemistry of Cells and Tissues written by Dieter Walz and published by Springer Science & Business Media. This book was released on 1995-10-30 with total page 328 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioelectrochemistry: Principles and Practice provides a comprehensive compilation of all the physicochemical aspects of the different biochemical and physiological processes. The role of electric and magnetic fields in biological systems forms the focus of this second volume in the Bioelectrochemistry series. The most prominent use of electric fields is found in some fish. These species generate fields of different strengths and patterns serving either as weapons, or for the purpose of location and communication. Electrical phenomena involved in signal transduction are discussed by means of two examples, namely excitation-contraction coupling in muscles and light transduction in photoreceptors. Also examined is the role of electrical potential differences in energy metabolism and its control. Temporal and spatial changes of the potential difference across the membranes of nerve cells are carefully evaluated, since they are the basis of the spreading and processing of information in the nervous system. The dielectric properties of cells and their responses to electric fields, such as electrophoresis and electrorotation, are dealt with in detail. Finally, the effects of magnetic fields on living systems and of low-frequency electromagnetic fields on cell metabolism are also considered. Further volumes will be added to the series, which is intended as a set of source books for graduate and postgraduate students as well as research workers at all levels in bioelectrochemistry.
Download or read book Mammalian Cell Viability written by Martin J. Stoddart and published by Humana Press. This book was released on 2011-04-12 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Whether the question is one of basic cell survival, or whether it is being used to correlate cell number to some other factor such as matrix synthesis, an estimate of cell viability is universally required. In Mammalian Cell Viability: Methods and Protocols, experts in the field describe methods from the most basic which can be performed in any laboratory, to some which require specific pieces of equipment. Initially focusing on methods for monolayer and suspension cells, later chapters describe methods for determining viability within tissue sections and 3 dimensional culture systems. Finally, methods requiring highly specialized equipment are described in order to explain what is possible. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting and avoiding known pitfalls. Practical and adaptable, Mammalian Cell Viability: Methods and Protocols serves as a self-contained laboratory manual useful to both experienced researchers and those new to this incredibly important and influential field.
Download or read book Biological Toxins and Bioterrorism written by P. Gopalakrishnakone and published by Springer. This book was released on 2015-01-22 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biological toxins are an important part of our world, a reality with which we need to cope, so in parallel with understanding their mechanisms of action and thereby improving our fundamental knowledge, there are successful efforts to utilize them as therapeutics against some debilitating human and animal diseases. In view of the complexity of different types of biotoxins and the broad range of toxin structure, physiology, utility, and countermeasures including regulatory issues, it was thus aimed to compile a book on biotoxins and bioweapons. This reference work in the Toxinology handbook series gathers together knowledge from around the globe about naturally inspired and manufactured biological weapons. The authors describe how they work; how authorities may detect their presence, prevent their use, and diagnose their impacts; and the means by which medical and paramedical professionals may treat victims. Also described are how they have been used to further our knowledge and what insights they have given us into evolutionary and physiological processes. Finally, it is also discussed how these toxins can be used as therapeutics and what the implications of such therapeutics are to their use as biothreat agents. This volume provides a reference accessible to scientists, educators, and medical experts alike with an interest in biotoxins, focusing on the major toxins used as bioweapons. Regulatory agencies will also benefit from the information provided in this book. Some in the intended audience may need to understand how they elicit their effects and how we can defend ourselves against them. Others may be interested in the sometimes colorful histories that surround this subset of biotoxins that can be and, in some cases, have been used as weapons.
