Drug Testing In Vitro: Breakthroughs and Trends in Cell Culture Technology Book

Foreword   Rolf Krebs     V
Preface     VII
List of Contributors     XVII
Emerging In-Vitro Culture Technologies     1
Intelligent Biomatrices and Engineered Tissue Constructs: In-Vitro Models for Drug Discovery and Toxicity Testing   Philip Lazarovici   Mengyan Li   Anat Perets   Mark J. Mondrinos   Shimon Lecht   Christopher D. Koharski   Paul R. Bidez III   Christine M. Finck   Peter I. Lelkes     3
Introduction     3
Intelligent Biomaterials and Scaffolds for Tissue Engineering     4
Synthetic Materials     4
Natural Biomaterials     5
Fabrication of Scaffolds for Tissue Engineering     7
Electrospinning     7
Controlled Lyophilization     9
Acellularization     10
Progress and Achievements in Liver Tissue Engineering     11
The Liver     11
Scaffolds for Liver Tissue Engineering     12
Pharmaceutical Applications of Tissue-Engineered Liver Models     15
Conclusions and Novel Trends in Liver Tissue Engineering     16
Cardiac Tissue Engineering: Cells and Models     16
Cardiac Tissue Engineering     16
Cells used in Cardiac Tissue Engineering     17
Culture Models of Cardiac Tissue-Engineered Constructs     18
Specific Scaffolds Developed for Cardiac Tissue Engineering     20
In-Vitro-Engineered Pulmonary Tissue Models: Progress and Challenges     21
Lung Tissue Engineering: The Current State of Play     21
Existing In-Vitro Pulmonary Cell and Tissue Culture Biological Models     26
Potential of Alveolar Tissue Models as Disease Models in Pharmaceutical Sciences     27
The Future: Toward Engineered 3D Alveolar Tissue for Cell Therapy and Pharmacological Models     27
In-Vitro Models of the Blood-Brain Barrier (BBB)     28
The BBB, a Neurovascular Physiological Unit: The Concept     28
In-Vitro BBB Models: Cells and Devices     32
BBB In-Vitro Models: From First to Third Generation; the Biological Approach     35
Trends in Tissue Engineering: Realistic In-Vitro BBB Pharmacological Models     36
Conclusions for BBB In-Vitro Models     38
References     39
An Overview on Bioreactor Design, Prototyping and Process Control for Reproducible Three-Dimensional Tissue Culture   Ralf Portner   Christoph Giese     53
Introduction     53
Important Aspects for Bioreactor Design      55
Culture Systems and Bioreactors Used in Tissue Engineering     57
The Operation of Bioreactors     59
3D Systems Used for Drug Testing     62
Modeling of Bioreactor Systems for Tissue Engineering     62
The Artificial Immune System     65
Matrices     68
Microenvironment     68
Monitoring     68
Conclusions     69
References     70
An Overview on Bioelectronic and Biosensoric Microstructures Supporting High-Content Screening in Cell Cultures   Andrea A. Robitzki   Andree Rothermel     79
The Potential of Drug Development and Demand on High-Content Screening Systems     79
Post-Genomics or Proteomics: An Analysis of Manifold Systems and Functional Monitoring of Drugs     79
Pharmaceutical Research and High-Technology Platforms in the Biohybrid Technology Field     80
Synergy of Microchip Technology and Living Cells     81
Microfabrication Techniques to Generate Miniaturized Chip Components     82
Microelectrode-Based Techniques for Analyzing Cellular Parameters: Possible Use of Real-Time and HTS of Drugs Without Labeling     85
Impedance Spectroscopy: Screening the Cellular Parameters of Electrophysiologically Inactive Cells     85
Intracellular Recording of Electroactive Cells: Chip-Based, Automated Patch-Clamp Recording     91
Extracellular Recording of Electrically Excitable Cells: Multiple Site Recording of Field Potentials by MEAs     93
Concluding Remarks: Secondary Screening for Safety and Cost-Effective Drug Testing and Discovery     96
References     97
Novel In-Vitro Exposure Techniques for Toxicity Testing and Biomonitoring of Airborne Contaminants   Amanda Hayes   Shahnaz Bakand   Chris Winder     103
Introduction     103
The Inhalation of Air Contaminants     103
Toxicological Assessment     105
In-Vitro Toxicological Studies     107
Applications of In-Vitro Test Methods     107
In-Vitro Toxicity Endpoints     108
In-Vitro Toxicity Testing of Air Contaminants     109
Indirect Methods     111
Direct Methods     112
Conclusions     115
References     116
Primary Tissues and Cell Lines in Drug Screening/Testing     125
Drug Screening Using Cell Lines: Cell Supply, High-Throughput and High-Content Assays   Christa Burger   Oliver Poschke   Mirek R. Jurzak     127
Introduction     127
Cell Lines for HTS      128
Selection of the Most Suitable Cell Line     128
Optimizing Cell Cultivation     130
Adherence     130
pH and Temperature     130
Media and Additives     131
Solvent Tolerance     131
Cell Density     131
Optimizing the Reproducibility of Seeding     132
Signal Shift     132
Edge Effect     132
Cell Production and Plate Delivery     132
The Amount of Cells Needed     132
Cell Storage     133
Conventional Cellular Screening Assays     134
General HTS Assay Prerequisites     134
Evaluation of Assay Quality     134
ELISA-Based Assays     135
Radiometric Cellular Assays     136
Reporter Gene Assays     137
Second Messenger Assays     138
Ion Channel Assays     138
The Definition of High-Content Screening     139
Instrumentation for HCS     139
Reagents (Fluorescent Probes) for HCS     140
Low-Molecular-Weight Fluorophores     140
Genetically Encoded Reporter for Fluorescence Detection     141
Assays and Target-Based Applications of HCS     142
GPCRs      142
Kinases     143
Other Drug Targets     144
HCS Applications Targeting Generic Cellular Parameters and Morphology     145
Outlook     146
References     147
Cell Lines and Primary Tissues for In-Vitro Evaluation of Vaccine Efficacy   Anthony Meager     153
Introduction     153
Measurement of Antigen Expression     155
Post-Vaccination Testing     158
Ex-Vivo Detection of Antigen-Specific T Cells     360
ELISPOT Assay     160
Cytokine Capture Assay and Intracellular Cytokine Staining     162
Measurement of T-Cell Cytotoxicity     163
Current Knowledge on T-Cell Responses in Vaccine Trials     165
Future Directions     167
References     168
Designer Cells Derived from Primary Tissue and Designed Cell Lines as a Sustainable Cell Source for Drug Discovery and Safety Assessment   Volker Sandig   Ingo Jordan     177
Introduction     177
Suitability and Limitations of Primary Cells as Physiologic Models     178
Tumor Cell Lines: Sometimes an Alternative     179
Immortalization by Design: Infinite Proliferation and a Differentiated Phenotype?      179
Telomerase: the Primary Target in Human Cells     179
Inactivation of Rb and p53 Pathways     181
Conditional Immortalization     183
Designed Cells in Complex Drug Tests     184
Cell Properties Required for Complex Screening Systems     184
Complex Designer Cells in Screens     185
Viruses and Host Cells in Drug Tests     189
Viruses and Designed Host Cells     190
Defined Viral and Cellular Pathways and Designed Host Cells     190
Virus Field Isolates and Designed Host Cells     192
Designed Viruses and Designed Host Cells     193
Designed Host Cells Combined     194
References     196
How Human Embryonic Stem Cell Research Can Impact In-Vitro Drug Screening Technologies of the Future   Andre Schrattenholz   Martina Klemm     205
Introduction     205
First Excursion: Protein Surrogate Biomarker Signatures     208
Second Excursion: Validation     211
Reproductive Toxicology and In-Vitro Tests     213
Reproductive Toxicology and hESC     214
Efficacy and Mode of Action Studies: Systems Biology Using Embryonic Stem Cell-Based Screening Systems     218
Conclusions and Outlook      221
References     222