Chien Bioengineering in Cell and Tissue Research

1. Auflage 2008
ISBN: 978-3-540-75409-1
Verlag: Springer-Verlag
Format: PDF
Kopierschutz: Adobe DRM (»Systemvoraussetzungen)

E-Book, Englisch, 693 Seiten

Cutting edge research in cell and tissue research abounds in this review of the latest technological developments in the area. The chapters are written by excellent scientists on advanced, frontier technology and address scientific questions that require considerable engineering brainpower. The aim is to provide students and scientists working in academia and industry new information on bioengineering in cell and tissue research to enhance their understanding and innovation.

Autoren/Hrsg.

Chien, Shu

Inhaltsverzeichnis

1;Editorial Board;5
2;Introduction;6
3;Contents;15
4;Part I Reporter Genes in Cell Based ultra High Throughput Screening;29
4.1;1 Reporter Genes in Cell Based ultra High Throughput Screening;29
4.1.1;1.1 Introduction;29
4.1.2;1.2 From Gene to Target;30
4.1.3;1.3 Screening Assay Classes;30
4.1.4;1.4 Reporter Gene Classes;31
4.1.5;1.5 Flash-Light Reporter Genes;32
4.1.6;1.6 Glow-Light Reporter Genes;34
4.1.7;1.7 Coelenterazine Dependent Luciferases;34
4.1.8;1.8 Luciferin Dependent Luciferases;36
4.1.9;1.9 Non-Luciferase Glow-Light Reporter Genes;37
4.1.10;1.10 Fluorescent Proteins;38
4.1.11;1.11 Cell Based Assay Formats in ultra High Throughput Screening (uHTS);40
4.1.12;1.12 Reporter Genes in uHTS;42
4.1.13;1.13 Photoprotein Readouts and Cell-Based Assay Development;43
4.1.14;1.14 Multiplexing Reporter Gene Readouts;44
4.1.15;1.15 Ultra High Throughput Screening;45
4.1.16;References;47
4.2;2 Gene Arrays for Gene Discovery;49
4.2.1;2.1 Introduction;49
4.2.2;2.2 Data Processing 2.2.1 Challenges in Data Acquisition and Processing;51
4.2.3;2.2.2 Data Preprocessing: An Overview;51
4.2.4;2.3 Gene Discovery by Gene Clustering;53
4.2.5;2.4 TGF-ß1 Signaling in Dendritic Cells Assessed by Gene Expression Profiling;55
4.2.6;2.4.1 Linking Gene Expression Data to Knowledge-based Databases;58
4.2.7;2.5 Conclusions;58
4.2.8;References;61
4.3;3 Physical Modulation of Cellular Information Networks;63
4.3.1;3.1 Introduction;63
4.3.2;3.2 Cellular Responses to Physical Stimulation;64
4.3.3;3.2.1 Electrical Potential;65
4.3.4;3.2.2 Hydrostatic Pressure;68
4.3.5;3.2.3 Shear Stress;69
4.3.6;3.2.4 Heat Shock;70
4.3.7;3.3 Electrically Controlled Proliferation Under Constant Potential Application 3.3.1 Electrical Potential- Controlled Cell Culture System;71
4.3.8;3.3.2 Cell Viability Under Constant Potential Application;71
4.3.9;3.3.3 Electrical Modulation of Cellular Proliferation Rate;73
4.3.10;3.4 Modulated Proliferation Under Extreme Hydrostatic Pressure;73
4.3.11;3.5 Electrically Modulated Gene Expression Under Alternative Potential Application 3.5.1 Electrically Stimulated Nerve Growth Factor Production;76
4.3.12;3.5.2 Electrically Induced Differentiation of PC12 Cells;78
4.3.13;3.6 Cellular Engineering to Enhance Responses to Physical Stimulation;80
4.3.14;3.7 Concluding Remarks;83
4.3.15;References;85
5;Part II Cell and Tissue Imaging;88
5.1;4 Fluorescence Live-Cell Imaging: Principles and Applications in Mechanobiology;89
5.1.1;4.1 Introduction;89
5.1.2;4.2 Fluorescence Proteins;90
5.1.3;4.3 Fluorescence Microscopy;93
5.1.4;4.4 Applications in Mechanobiology;95
5.1.5;4.5 Perspective in Cardiovascular Physiology and Diseases;102
5.1.6;References;104
5.2;5 Optical Coherence Tomography (OCT) – An Emerging Technology for Three- Dimensional Imaging of Biological Tissues;108
5.2.1;5.1 Introduction;108
5.2.2;5.2 Optical Coherence Tomography (OCT);110
5.2.3;5.3 Applications in Tissue Engineering;118
5.2.4;5.4 Conclusion;121
5.2.5;References;122
5.3;6 Ultrasonic Strain Imaging and Reconstructive Elastography for Biological Tissue;126
5.3.1;6.1 Introduction;126
5.3.2;6.2 Ultrasound Elastography;128
5.3.3;6.3 Reconstructive Ultrasound Elastography;137
5.3.4;6.4 Medical Results of Elastography;146
5.3.5;6.5 Results of an Intravascular Ultrasound Study;147
5.3.6;6.6 Summary and Conclusion;150
5.3.7;References;152
6;Part III Regenerative Medicine and Nanoengineering;156
6.1;7 Aspects of Embryonic Stem Cell Derived Somatic Cell Therapy of Degenerative Diseases;157
6.1.1;7.1 Introduction;157
6.1.2;7.2 Rationale for the Cardiac Tissue Engineering;158
6.1.3;7.3 Embryonic Stem Cells as an Unlimited Source for Cardiomyocytes;159
6.1.4;7.4 Therapeutical Cloning of Embryonic Stem Cells;162
6.1.5;7.5 Stem Cell Derived Cardiomyocytes;165
6.1.6;7.6 Cardiac Tissue Slices;167
6.1.7;7.7 Bioartificial Heart Tissue Based on Biomaterials;169
6.1.8;7.8 Scaffolds for Cardiac Tissue Engineering;170
6.1.9;7.9 The Ideal Cell;173
6.1.10;7.10 Preparation of Cells for In-Vitro Tissue Engineering: Cell Permeable Cre/ loxP System;174
6.1.11;7.11 Outlook;176
6.1.12;References;178
6.2;8 Collagen Fabrication for the Cell-based Implants in Regenerative Medicine;180
6.2.1;8.1 Regenerative Medicine;180
6.2.2;8.2 The Cell-Based Implants;181
6.2.3;8.3 Requirements of Materials for the Cell-Based Implants;182
6.2.4;8.4 Biomaterials in the Cell-Hybridization;183
6.2.5;8.5 Characteristics of Collagen;184
6.2.6;8.6 Fabrication of Collagen;186
6.2.7;8.7 Collagen in the Cell-based Implants;191
6.2.8;8.7.1 Skin Regeneration;191
6.2.9;8.7.2 Bone Reconstruction;196
6.2.10;8.7.3 Esophagus Replacement;198
6.2.11;8.7.4 Wound Healing promoting Anti-Adhesive Matrix;203
6.2.12;8.7.5 Liver Regeneration;205
6.2.13;8.8 Discussion;208
6.2.14;References;211
6.3;9 Tissue Engineering – Combining Cells and Biomaterials into Functional Tissues;213
6.3.1;9.1 Introduction;213
6.3.2;9.2 The Cells;214
6.3.3;9.3 The Material;222
6.3.4;References;229
6.4;10 Micro and Nano Patterning for Cell and Tissue Engineering;235
6.4.1;10.1 Overview;235
6.4.2;10.2 Regulation of Cell Functions by Matrix Patterning;236
6.4.3;10.3 Topographic Regulation of Cell Functions;241
6.4.4;10.4 Engineering 3D Environments with Micro Features;242
6.4.5;10.5 Nano Patterning for Cell and Tissue Engineering;245
6.4.6;10.6 Perspective;247
6.4.7;References;248
6.5;11 Integrative Nanobioengineering: Novel Bioelectronic Tools for Real Time Pharmaceutical High Content Screening in Living Cells and Tissues;249
6.5.1;11.1 Introduction;249
6.5.2;11.2 Real Time Monitoring and High Content Screening;250
6.5.3;11.3 Outlook and Future Aspects;263
6.5.4;References;264
7;Part IV Soft Materials in Technology and Biology – Characteristics, Properties, and Parameter Identification;268
7.1;12 Soft Materials in Technology and Biology – Characteristics, Properties, and Parameter Identification;268
7.1.1;12.1 Introduction;268
7.1.2;12.2 Material Description ;270
7.1.3;12.3 Basics of Continuum Mechanics;287
7.1.4;12.4 Basics of Material Theory;293
7.1.5;12.5 Material Laws for Technical and Biological Polymers;309
7.1.6;12.6 Volume Change in Biopolymers;318
7.1.7;12.7 Summary and Outlook;325
7.1.8;References;327
7.2;13 Modeling Cellular Adaptation to Mechanical Stress;330
7.2.1;13.1 Introduction;330
7.2.2;13.2 A Brief Review of Stretch-Induced Cell Remodeling;331
7.2.3;13.3 Measurements, Modeling, and Mechanotransduction;335
7.2.4;13.4 A New Approach for the Study of the Mechanobiology of Cell Stretching;345
7.2.5;13.5 Illustrative Examples;354
7.2.6;13.6 Closure;357
7.2.7;References;359
7.3;14 How Strong is the Beating of Cardiac Myocytes? – The CellDrum Solution;362
7.3.1;14.1 Introduction;362
7.3.2;14.2 The CellDrum Technique;365
7.3.3;14.3 Preparation of Samples;367
7.3.4;References;378
7.4;15 Mechanical Homeostasis of Cardiovascular Tissue;381
7.4.1;15.1 Introduction;381
7.4.2;15.2 Shear Stress and Scaling Laws of Vascular System;382
7.4.3;15.3 Stress and Strain ;384
7.4.4;15.4 Intramural Stress and Strain;386
7.4.5;15.5 Perturbation of Mechanical Homeostasis;388
7.4.6;15.6 Limitations, Implications and Future Directions;394
7.4.7;References;397
7.5;16 The Role of Macromolecules in Stabilization and De- Stabilization of Biofluids;402
7.5.1;16.1 Introduction;402
7.5.2;16.2 The Effects of Macromolecules on the Stability of Colloids;405
7.5.3;16.3 Macromolecular Depletion at Biological Interfaces;406
7.5.4;16.4 Cell–Cell Interactions Mediated by Macromolecular Depletion;410
7.5.5;16.5 Stabilization of Bio-Fluids via Macromolecules;415
7.5.6;16.6 Destabilization of Bio-Fluids via Macromolecular Binding;417
7.5.7;16.7 Conclusion & Outlook;418
7.5.8;References;420
7.6;17 Hemoglobin Senses Body Temperature;424
7.6.1;17.1 Instead of an Introduction;424
7.6.2;17.2 Physiological Aspects of Thermoregulation in the Body;427
7.6.3;17.3 Red Blood Cells;429
7.6.4;17.4 Temperature Transition in RBC Passage Through Micropipettes;429
7.6.5;17.5 The Molecular Mechanism of the Micropipette Passage Transition;430
7.6.6;17.6 Hemoglobin Viscosity Transition;432
7.6.7;17.7 Circular Dichroism Transition in Diluted Hb Solutions;433
7.6.8;17.8 A RBC Volume Transition Revealed with Micropipette Studies;435
7.6.9;17.9 Micropipette Passage Transition in D2O Buffer;437
7.6.10;17.10 NMR T1 Relaxation Time Transition of RBCs in Autologous Plasma;438
7.6.11;17.11 Colloid Osmotic Pressure Transition of RBC Suspended in Plasma;441
7.6.12;17.12 The Temperature Transition Effect so Far;442
7.6.13;17.13 Strange coevals – Ornithorhynchus anatinus and Tachyglossus aculeatus;443
7.6.14;17.14 Hb Temperature Transition of Species with Body Temperatures Different from 37 C;444
7.6.15;17.15 Molecular Structural Mechanism of the Temperature Transitions;447
7.6.16;17.16 Physics Meets Physiology;449
7.6.17;References;452
8;Part V Bioengineering in Clinical Applications;457
8.1;18 Nitric Oxide in the Vascular System: Meet a Challenge;458
8.1.1;18.1 Nitric Oxide: NO;458
8.1.2;18.2 NO in Vascular Biology;458
8.1.3;18.3 Key Questions;460
8.1.4;18.4 Assessment of NO Mediated Vasoactivity;460
8.1.5;18.5 From the In-Vivo and Ex-Vivo Detection of NO Effects to Biochemical Assessment of NO;462
8.1.6;18.6 On the Road to a Potential Sensitive Marker for NO Formation: Is Nitrite a Candidate?;463
8.1.7;18.7 More Information About NO Interactions in the Blood;465
8.1.8;18.8 Intravascular Sources of NO;466
8.1.9;18.9 The Potential Relevance of RBC NOS Activity;466
8.1.10;18.10 Outlook;469
8.1.11;References;472
8.2;19 Vascular Endothelial Responses to Disturbed Flow: Pathologic Implications for Atherosclerosis;476
8.2.1;19.1 Introduction;476
8.2.2;19.2 Endothelial Dysfunction is a Marker of Atherosclerotic Risk;477
8.2.3;19.3 Correlation Between Lesion Locations and Disturbed Flow Regions of the Arterial Tree;478
8.2.4;19.4 In Vitro Studies on the Effects of Disturbed Flow on ECs;480
8.2.5;19.5 In Vivo Studies on the Effects of Disturbed Flow on ECs;490
8.2.6;19.6 Summary and Conclusions;492
8.2.7;References;495
8.3;20 Why is Sepsis an Ongoing Clinical Challenge? Lipopolysaccharide Effects on Red Blood Cell Volume;503
8.3.1;20.1 Introduction;504
8.3.2;20.2 Physiopathological Events During Sepsis;505
8.3.3;20.3 Markers in Clinical Diagnosis of Sepsis;505
8.3.4;20.4 Microcirculation and Sepsis;506
8.3.5;20.5 Therapy;506
8.3.6;20.6 Activated Protein C;507
8.3.7;20.7 Red Blood Cell Behaviour During Sepsis;507
8.3.8;20.8 New Perspective;508
8.3.9;References;513
8.4;21 Bioengineering of Inflammation and Cell Activation: Autodigestion in Shock;515
8.4.1;21.1 Introduction;516
8.4.2;21.2 Inflammation in Shock and Multi-Organ Failure;517
8.4.3;21.3 The Pancreas as a Source of Cellular Activating Factors and the Role of Serine Proteases;518
8.4.4;21.4 Blockade on Pancreatic Digestive Enzymes in the Lumen of the Intestine;519
8.4.5;21.5 What Mechanisms Prevent Auto-digestion?;521
8.4.6;21.6 Triggers of Shock Increase Intestinal Wall Permeability;521
8.4.7;21.7 Intestine as Source of Inflammatory Mediators in Shock;522
8.4.8;21.8 Characterization of Protease-Derived Shock Factors;522
8.4.9;21.9 Cytotoxic Factors Derived from the Intestine;524
8.4.10;21.10 Removal or Blockade of Intestinal Cytotoxic Mediators;525
8.4.11;21.11 Conclusions;526
8.4.12;References;528
8.5;22 Percutaneous Vertebroplasty: A Review of Two Intraoperative Complications;531
8.5.1;22.1 Introduction;531
8.5.2;22.2 Vertebroplasty: Minimally Invasive and Cost- Effective Solution;532
8.5.3;22.3 Extravertebral Biomechanics: Excessive Delivery Pressure;533
8.5.4;22.4 Intravertebral Biomechanics: Risk of Extravasation;535
8.5.5;22.5 Injectable Biomaterials;537
8.5.6;22.6 Discussion;538
8.5.7;References;540
9;Part VI Plant and Microbial Bioengineering;543
9.1;23 Molecular Crowding: AWay to Deal with Crowding in Photosynthetic Membranes;544
9.1.1;23.1 In the Crowd;544
9.1.2;23.2 Macromolecular Crowding;546
9.1.3;23.3 Photosynthesis in a Crowded Environment;553
9.1.4;23.4 Crowding Effects in Photosynthetic Membranes;559
9.1.5;23.5 Summary and Outlook;571
9.1.6;References;575
9.2;24 Higher Plants as Bioreactors. Gene Technology with C3- Type Plants to Optimize CO2 Fixation for Production of Biomass and Bio- Energy;580
9.2.1;24.1 Introduction;580
9.2.2;24.2 The C3 and C4 CO2 Fixation Mechanisms;582
9.2.3;24.3 The Metabolism of Glycolate in Escherichia coli and in Some Green Algae;586
9.2.4;24.4 Increased Biomass Production in Transgenic Arabidopsis Plants Containing the E. coli Glycolate Pathway in the Chloroplasts 24.4.1 The Strategy to Improve CO2 Fixation and Hence Photosynthesis;588
9.2.5;24.4.2 Establishment of the E. coli Glycolate Pathway in Arabidopsis Chloroplasts;589
9.2.6;24.5 Analysis of the GT-DEF Transgenic Plants. DNA, RNA, Proteins, Physiology, Growth and Production of Biomass;593
9.2.7;24.6 Summary;596
9.2.8;References;598
9.3;25 Controlling Microbial Adhesion: A Surface Engineering Approach;600
9.3.1;25.1 The LostWorld of Sessile Microorganisms;600
9.3.2;25.2 Biotechnological Potential of Adhered Microorganisms and Its Limitations;603
9.3.3;25.3 Physicochemical Aspects of Microbial Adhesion;606
9.3.4;25.4 Biological Aspects of Microbial Adhesion;608
9.3.5;25.5 Surface Conditioning as a Tool FacilitatingMicrobial Adhesion;611
9.3.6;References;620
9.4;26 Air Purification Technology by Means of Cluster Ions Generated by Plasma Discharge at Atmospheric Pressure;623
9.4.1;26.1 Ion Generating Device;623
9.4.2;26.2 Characteristics of Positive and Negative Ions;624
9.4.3;26.3 Effect of Removing Airborne Bacteria;625
9.4.4;26.4 Effect of Removing Floating Fungi (Mould);628
9.4.5;26.5 Effect of Deactivating Floating Viruses;629
9.4.6;26.6 Virus Deactivation Model Using Cluster Ions;631
9.4.7;26.7 Allergen Deactivation Effect;632
9.4.8;26.8 Conclusion;635
9.4.9;References;636
9.5;27 Astrobiology;637
9.5.1;27.1 Introduction;637
9.5.2;27.2 Origin and History of Life on Earth;638
9.5.3;27.3 Impact Scenario and Interplanetary Transport of Life;642
9.5.4;27.4 Strategies of Life to Adapt to Extreme Environments;643
9.5.5;27.5 Signatures of Life;645
9.5.6;27.6 Criteria for Habitability;647
9.5.7;27.7 Planets and Moons in Our Solar System That are of Interest to Astrobiology;649
9.5.8;27.8 Planetary Protection;654
9.5.9;27.9 Search for Life Beyond the Solar System;657
9.5.10;27.10 Outlook;658
9.5.11;References;660
10;Authors;662
11;Index;676

Leseproben

Chapter 1 Reporter Genes in Cell Based ultra High Throughput Screening (p. 3-4)

Stefan Golz

Bayer Healthcare AG, Institute for Target Research, 42096Wuppertal, Germany, [e-mail]

Abstract Pharma research in most organizations is organized in discrete phases together building a "value chain" along which discovery programs process to fi- nally drug candidates for clinical testing. The process envisioned to identify targetspecific modulators lacking several side effects. Following a technical assessment of the targets "drugability", the probability to identify small molecule modulators, and technical feasibility target-specific assays are developed to probe the corporate compound collection for meanful leads. "High-Throughput-Screening" (HTS) started roughly one decade ago with the introduction of laboratory automation to handle the different assay steps typically performed in microtiter plates. Today a large arsenal of screening technologies is available for researchers in industry and academia to set up uHTS or HTS assays. Here the use of reporter genes offer an alternative for following signal transduction pathways from receptors at the cell surface to nuclear gene transcription in living cells.

1.1 Introduction

The modern drug research process has reversed the classical pharmacological strategy. Today, research programs are initiated based on biological evidence suggesting a particular gene or gene product to be a meaningful target for small molecule drugs useful for therapies. The process envisioned to identify target-specific modulators lacking several side effects. Also, it allows setting up a linear drug discovery process starting from target identification to finally delivering molecules for clinical development. One central element is lead discovery through high-throughput screening of comprehensive corporate compound collections. Pharma research in most organizations is organized in discrete phases together building a "value chain" along which discovery programs process to finally drug candidated for clinical testing (Hüser et al. 2006).

This pipeline is fueled by targets suggested from external or in-house generated data suggesting a gene or gene product to be disease relevant. Today a large arsenal of technologies is available for researchers in industry and academia to generate data in support of a functional link between a given gene and a disease state.

1.2 From Gene to Target

Following a technical assessment of the targets "drugability" (Hopkins and Groom, 2002), the probability to identify small molecule modulators, and technical feasibility target-specific assays are developed to probe the corporate compound collection for meanful leads. Lead discovery in the pharmaceutical industry today still depends largely on experimental screening of compound collections. To this end, the industry has invested heavily in expanding their compound files and established appropiate screening capabilities to handle large numbers of compounds within a reasonable period of time. "High-Troughput-Screening" (HTS) started roughly one decade ago with the introduction of laboratory automation to handle the different assay steps typically performed in microtiter plates. HTS technologies during the last decade have witnessed remarkable developments. Assay technologies have advanced to provide a large variety of various cell-based and biochemical test formats for a large spectrum of disease relevant target classes (Walters and Namchuk, 2003). In parallel, further miniaturization of assays volumes and parallelization of processing have further increased the test throughput. The ultra-high-throughput is required to fully exploit big compound files of >,1million compounds and is performed entirely in 1536-well plates with assay volumes between 5 – 10 ìl. This assay carrier together with fully-automated robotic systems allow for testing in excess of 200,000 compounds per day. The comprehensive substance collection, together with sophisticated screening technologies, have resulted in a clear advantages in lead discovery especially for poorly druggable targets with a poor track record in the past.