Methods in bioengineering : alternatives to animal testing /

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 e...

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Detalles Bibliográficos
Clasificación:Libro Electrónico
Otros Autores: Maguire, Timothy J. (Timothy John), Novik, Eric
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Boston, Mass. : Artech House, ©2010.
Colección:Artech House methods in bioengineering series.
Temas:
Animal experimentation > Moral and ethical aspects.
Animal welfare > Moral and ethical aspects.
Human-animal relationships > Moral and ethical aspects.
Expérimentation animale > Aspect moral.
Animaux > Protection > Aspect moral.
Relations homme-animal > Aspect moral.
PHILOSOPHY > Ethics & Moral Philosophy.
Acceso en línea:Texto completo
Tabla de Contenidos:
  • 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.

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