Title Methods in bioengineering : alternatives to animal testing / Tim Maguire, Eric Novik, editors

Published Boston, Mass. : Artech House, ©2010

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Description 1 online resource (xiii, 283 pages) : illustrations (some color) Series Methods in bioengineering series Artech House methods in bioengineering series. Contents Preface -- Chapter 1 Current Methods for Prediction of Human Hepatic Clearance Using In Vitro Intrinsic Clearance -- 1.1 Introduction -- 1.2 Materials -- 1.3 Methods -- 1.3.1 Thawing the hepatocytes -- 1.3.2 Clearance study using a hepatocyte suspension -- 1.3.3 Clearance study using a plated hepatocyte culture -- 1.3.4 Clearance study using a plated hepatocyte culture under a flow condition -- 1.3.5 Sampling for the clearance study -- 1.3.6 Sample analysis using LC-MS/MS -- 1.4 Data Acquisition, Anticipated Results, and Interpretation -- 1.4.1 Hepatocyte suspension and plated hepatocyte system -- 1.4.2 Physiologically based microfluidic systems -- 1.5 Discussion and Commentary -- 1.5.1 Hepatocyte suspension system -- 1.5.2 Plated hepatocyte system -- 1.5.3 Physiologically based microfluidic systems -- 1.6 Summary -- References -- Chapter 2 Use of Permeability from Cultured Cell Linesand PAMPA System and Absorption from Experimental Animals for the Prediction of Absorption in Humans -- 2.1 Introduction -- 2.2 Materials -- 2.3 Methods -- 2.3.1 Cultured cell system -- 2.3.2 PAMPA system -- 2.3.3 In vivo absorption measurements -- 2.4 Data Acquisition, Anticipated Results, and Interpretation -- 2.4.1 Data analysis -- 2.4.2 Results and interpretation -- 2.5 Discussion and Commentary -- 2.5.1 Cell culture and PAMPA systems -- 2.5.2 Absorption in experimental animals -- 2.5.3 Rats -- 2.5.4 Dogs -- 2.5.5 Monkeys -- 2.6 Summary -- References -- Chapter 3 Aggregating Brain Cell Cultures for Neurotoxicity Tests -- 3.1 Introduction -- 3.2 Experimental Design -- 3.3 Materials -- 3.3.1 Animals -- 3.3.2 Special equipment -- 3.3.3 Reagents -- 3.3.4 Preparation of solutions and media -- 3.4 Methods -- 3.4.1 Washing and sterilizing the glassware -- 3.4.2 Cell isolation and culture preparation 3.4.3 Maintenance of aggregating brain cell cultures (media replenishmentand subdivision) -- 3.4.4 Preparation and treatment of replicate cultures -- 3.4.5 Harvest of replicate cultures for various analytical procedures -- 3.4.6 Examples of sample preparation and use for various analytical procedures -- 3.4.7 Data Analysis -- 3.5 Anticipated Results -- 3.6 Discussion and Commentary -- 3.7 Application Notes -- 3.8 Summary Points -- Acknowledgments -- References -- Chapter 4 Approaches Towards a Multiscale Model of Systemic Inflammation in Humans -- 4.1 Introduction -- 4.2 Materials -- 4.2.1 Human endotoxin model and data collection -- 4.3 Methods -- 4.3.1 Transcriptional dynamics and intrinsic responses -- 4.3.2 Modeling inflammation at the cellular level -- 4.3.3 Modeling inflammation at the systemic level -- 4.4 Results -- 4.4.1 Elements of the multiscale host response model of human inflammation -- 4.4.2 Estimation of relevant model parameters -- 4.4.3 Qualitative assessment of the model -- 4.5 Conclusions -- Acknowledgments -- References -- Chapter 5 A Liposome Assay for Evaluating the Ocular Toxicity of Chemicals -- 5.1 Introduction -- 5.2 Experimental Design -- 5.3 Materials -- 5.4 Methods -- 5.4.1 Preparation of calcein-loaded liposomes -- 5.4.2 Separation of bulk calcein from loaded liposomes with Sephadex -- 5.4.3 Ocular toxicity experiments using dye-loaded liposomes -- 5.5 Data Acquisition, Anticipated Results, and Interpretation -- 5.6 Discussion and Commentary -- 5.7 Application Notes -- 5.8 Summary Points -- References -- Chapter 6 Prediction of Potential Drug Myelotoxicity by In Vitro Assays on Hematopoietic Progenitors -- 6.1 Introduction -- 6.2 Experimental Design -- 6.3 Materials -- 6.3.1 Reagents -- 6.4 Methods -- 6.4.1 Preparation of methylcellulose stocks -- 6.4.2 Source of murine hematopoietic progenitors 6.4.3 Source of human hematopoietic progenitors -- 6.4.4 Technical procedure for GM-CFU test -- 6.4.5 Passing from screening phase to IC determination phase -- 6.4.6 Incubator humidity test -- 6.4.7 Scoring the colonies -- 6.4.8 Criteria for colony counting -- 6.5 Data Acquisition, Anticipated Results, and Interpretation -- 6.5.1 Statistical guidelines -- 6.6 Discussion and Commentary -- 6.7 Application Notes -- 6.8 Summary Points -- Acknowledgments -- References -- Chapter 7 Epigenetically Stabilized Primary Hepatocyte Cultures: A Potential Sensitive Screening Toolfor Nongenotoxic Carcinogenicity -- 7.1 Introduction -- 7.2 Experimental Design -- 7.3 Materials -- 7.3.1 Reagents -- 7.3.2 Facilities/Equipment -- 7.4 Methods -- 7.4.1 Isolation of hepatocytes from rat liver -- 7.4.2 Cultivation of primary rat hepatocytes (Troubleshooting Table) -- 7.5 Data Acquisition -- 7.6 Anticipated Results and Interpretation -- 7.7 Discussion and Commentary -- 7.8 Application Notes -- 7.9 Summary Points -- Acknowledgements -- References -- Chapter 8 A Statistical Method to Reduce In Vivo Product Testing Using Related In Vitro Tests and ROC Analysis -- 8.1 Introduction -- 8.2 Experimental Design -- 8.3 Materials -- 8.4 Methods -- 8.4.1 Step-by-step protocol for the analysis of data using Analyse-It -- 8.5 Results -- 8.6 Discussion and Commentary -- 8.6.1 Selecting the proper secondary test -- 8.6.2 Determining the sample size for calibration and recalibration -- 8.6.3 Regulatory concerns -- 8.6.4 Determining the frequency of recalibration -- 8.6.5 Determining the need for confirmatory testing -- 8.6.6 Statistical analysis -- 8.7 Summary Points -- Acknowledgments -- References -- Chapter 9 Application of the Benchmark Approach in theCorrelation of In Vitro and In Vivo Data inDevelopmental Toxicity -- 9.1 Introduction -- 9.2 Materials and Methods 9.2.1 Derivation of in vitro BMC and BMD values -- 9.2.2 In vitro-in vivo correlation -- 9.3 Discussion and Commentary -- References -- Chapter 10 Three-Dimensional Cell Culture of Canine Uterine Glands -- 10.1 Introduction -- 10.2 Materials -- 10.2.1 Cell culture -- 10.2.2 Histological preparation for light microscopy -- 10.2.3 Histological preparation for electron microscopy -- 10.3 Methods -- 10.3.1 Cell culture -- 10.3.2 Histological preparation for light microscopy -- 10.3.3 Histological preparation for electron microscopy -- 10.3.4 Imaging -- 10.4 Anticipated Results -- 10.5 Discussion and Commentary -- 10.6 Application Notes -- 10.7 Summary Points -- References -- Chapter 11 Markers for an In Vitro Skin Substitute -- 11.1 Introduction -- 11.2 Experimental Design -- 11.3 Materials -- 11.3.1 Human tissue-engineered skin substitute reconstructed by the self-assembly approach -- 11.4 Methods -- 11.4.1 Preparation of solutions and materials for the in vitro fabrication of human skin substitutes by the self-assembly approach -- 11.4.2 In vitro fabrication of human skin substitutes by the self-assembly approach -- 11.4.3 Tissue preservation and sectioning -- 11.4.4 Preparation of solutions and materials for immunofluorescence -- 11.4.5 Immunofluorescent labeling of human skin substitutes -- 11.4.6 Histological analysis -- 11.4.7 Transmission electron microscopy -- 11.4.8 Statistical analysis -- 11.5 Anticipated Results -- 11.6 Discussion and Commentary -- 11.7 Application Notes -- 11.8 Summary Points -- Acknowledgments -- References -- Chapter 12 3D Culture of Primary Chondrocytes, Cartilage, and Bone/Cartilage Explants in Simulated Microgravity -- 12.1 Introduction -- 12.2 Experimental Design -- 12.2.1 Culture models -- 12.2.2 The RCCS bioreactor and its operational conditions -- 12.2.3 Animals -- 12.3 Materials 12.3.1 Equipment for cell/tissue culture and preparation of samples -- 12.3.2 Chemicals -- 12.4 Methods -- 12.4.1 Preparation of tissue explants -- 12.4.2 Isolation of chondrocytes -- 12.4.3 2D culture of isolated chondrocytes (traditional monolayer in staticfluid conditions) -- 12.4.4 3D culture of isolated chondrocytes (homotypic aggregates) -- 12.4.5 3D culture of fragments of articular cartilage explants -- 12.4.6 3D culture of undissected, complete proximal tibial epiphyses -- 12.4.7 Histomorphological study of chondrocytes and cartilage tissue -- 12.5 Anticipated Results -- 12.6 Discussion -- 12.6.1 Discussion of pitfalls -- 12.6.2 General discussion and commentary -- 12.7 Application Notes -- 12.8 Summary Points -- Acknowledgments -- References -- Chapter 13 Alternatives for Absorption Testing -- 13.1 Introduction -- 13.2 Materials -- 13.2.1 Franz diffusion cell -- 13.2.2 Consumables -- 13.2.3 Chemicals and solutions -- 13.2.4 Technical equipment -- 13.3 Methods -- 13.3.1 Skin preparation -- 13.3.2 Determination of skin penetration using the Franz cell setup -- 13.3.3 Determination of skin permeation using the Franz cell setup -- 13.3.4 Skin absorption studies with commercially available 3D skin models -- 13.3.5 Quality control -- 13.3.6 Data evaluation -- 13.3.7 Biostatistics -- 13.4 Results and Discussion -- 13.5 Discussion of Pitfalls and Troubleshooting -- 13.6 Summary -- References -- Chapter 14 A 3D Model of the Human Epithelial Airway Barrier -- 14.1 Introduction -- 14.2 Experimental Design -- 14.3 Materials -- 14.3.1 General materials -- 14.3.2 Epithelial cell cultures-thawing -- 14.3.3 Epithelial cell cultures-culturing -- 14.3.4 Isolation of monocyte-derived macrophages (MDM) and dendriticcells (MDDC) -- 14.3.5 Triple cell coculture -- 14.3.6 Transepithelial electrical resistance (TEER) measurements -- 14.4 Methods Summary Written and edited by recognized experts in the field, the new Artech House Methods in Bioengineering book series offers detailed guidance on authoritative methods for addressing specific bioengineering challenges. Offering a highly practical presentation of each topic, each book provides research engineers, scientists, and students with step-by-step procedures, clear examples, and effective ways to overcome problems that may be encountered. Providing alternatives to animal testing is one of the hottest topics in biomedical research, and this groundbreaking volume addresses this critical issues head on. This unique book presents techniques and methods at the forefront of scientific research that have the potential to replace certain whole animal tests. Moreover, this book provides a platform where other widely accepted techniques and scientific advancements can be collated into a concise set of methods that can be implemented within both academic and industrial communities. Publisher abstract Bibliography Includes bibliographical references and index Notes Print version record Subject Animal experimentation -- Moral and ethical aspects Animal welfare -- Moral and ethical aspects. Human-animal relationships -- Moral and ethical aspects PHILOSOPHY -- Ethics & Moral Philosophy. Animal experimentation -- Moral and ethical aspects Animal welfare -- Moral and ethical aspects Human-animal relationships -- Moral and ethical aspects Form Electronic book Author Maguire, Timothy J. (Timothy John) Novik, Eric. ISBN 9781608070121 1608070123

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