Tissue Engineering: A Primer with Laboratory Demonstrations

✍ Scribed by Jeong-Yeol Yoon

• Publisher: Springer
• Year: 2021
• Tongue: English
• Leaves: 274
• Edition: 1
• Category: Library

✦ Synopsis

Tissue Engineering: A Primer with Laboratory Demonstrations concisely covers the fundamental basics of tissue engineering. A series of simple, low-cost, and easy-to-implement laboratory modules are included in each chapter, along with experimental results with actual images and data, and a set of questions and discussion topics for each laboratory exercise. The textbook is appropriate for upper-undergraduate and graduate-level courses in cell and tissue engineering. The inclusion of images and data for all laboratory exercises also makes the book a valuable tool for scientists and engineers to learn the concepts in a hands-on and visual manner and lay a foundation to build their experiments towards their research and commercial development.

✦ Table of Contents

Preface
Contents
About the Author
Chapter 1: Introduction
1.1 Narrow Definition of Tissue Engineering
1.2 Early Attempt in Scaffold Development: Decellularized Matrix
1.3 Simple TE Transplant Example: Skin
1.4 Simple TE Transplant Example: Pancreas
1.5 Expanded Definition of Tissue Engineering: Organ-on-a-Chip (OOC)
1.6 Overview of This Book
References
Chapter 2: Cell Culture
2.1 What Is Cell Culture?
2.2 Cell Physiology: Cell Membrane and Cytoskeleton
2.3 Focal Adhesion
2.4 Cell Classification: Anchorage-Dependent Versus Anchorage-Independent Cells
2.5 Cell Classification: Normal Versus Immortalized Cells
2.6 Cell Classification: Normal Versus Stem Cells
2.7 Maintaining Sterile Environment: Biosafety Cabinet
2.8 Maintaining Sterile Environment: Autoclave
2.9 Cell Culture: CO2 Incubator
2.10 Cell Imaging: Fluorescence Microscope
2.11 Laboratory Task 1: Bacterial Cell Culture
2.12 Laboratory Task 2: Fluorescence Microscopic Imaging of Mammalian Cells
Reference
Chapter 3: Cell Metabolism
3.1 What Is Cell Metabolism?
3.2 Energy Currency: ATP
3.3 Cell Metabolism: Glycolysis and TCA Cycle
3.4 Glucose Transport and Uptake
3.5 Cell Metabolism: Glutaminolysis and TCA Cycle
3.6 Glutamine Transport and Uptake
3.7 Role of Oxygen in Cell Metabolism
3.8 Culture Media
3.9 Cell Feeding
3.10 Cell Passaging
3.11 Cell Growth Kinetics: First-Order Growth Model
3.12 Laboratory Task 1: Media Preparation and Cell Feeding
3.13 Laboratory Task 2: Cell Passaging
Reference
Chapter 4: Cell Imaging
4.1 Overview of Fluorescence Microscopy
4.2 Fluorescence
4.3 Fluorescent Dyes
4.4 Bioreceptors
4.5 Fluorescence Microscope
4.6 Photobleaching
4.7 Smartphone Fluorescence Microscope
4.8 Laboratory Task 1: Fluorescence Imaging of Nucleus and Cytoskeleton
4.9 Laboratory Task 2 (Alternative): Fluorescence Imaging of Nucleus, Cytoskeleton, and Focal Adhesion
References
Chapter 5: Stem Cells
5.1 What Are Stem Cells?
5.2 Why Do We Need Stem Cells for Tissue Engineering?
5.3 Embryonic Versus Adult Stem Cells
5.4 Use of Embryonic Stem Cells for Tissue Engineering Applications
5.5 Induced Pluripotent Stem Cells (iPSCs)
5.6 Isolation of Stem Cells
5.7 Culturing Stem Cells
5.8 Morphogenetic Factors
5.9 Hazards of Stem Cell Differentiation
5.10 Laboratory Task 1: Culturing Stem Cells
5.11 Laboratory Task 2: Embryoid Body Formation
References
Chapter 6: Biomaterial Surfaces
6.1 Development of Biomaterial Surfaces for Tissue Engineering
6.2 Size and Shape Requirements
6.3 Synthetic Materials
6.4 Natural Materials
6.5 Nonspecific Cell–Surface Interactions
6.6 Specific Cell–Surface Interactions
6.7 Bone Biomaterials, Apatite, and Bioglass
6.8 Hydrogels
6.9 Biodegradable Scaffolds
6.10 Encapsulation Scaffolds
6.11 Laboratory Task 1: Preparation of Various Biomaterial Surfaces
Objective 1. Preparation of PMMA Surface
Objective 2. Preparation of PDMS Surface
Objective 3. Smear Coating of Rat Tail Collagen Type I
6.12 Laboratory Task 2: Contact Angle Measurements
6.13 Laboratory Task 3: Surface Roughness Measurements
References
Chapter 7: Focal Adhesion
7.1 What Is Focal Adhesion?
7.2 Cell Adhesion and Proliferation on Biomaterial Surfaces
7.3 Cell Migration After Focal Adhesion
7.4 Morphogenesis
7.5 Laboratory Task 1: Fluorescence Staining of Focal Adhesion
Objective 1. Kill, Fix, and Perforate Cells
Objective 2. Anti-Vinculin Treatment
Objective 3. Anti-IgG-FITC and Phalloidin-TRITC
Objective 4. DAPI (Optional)
7.6 Laboratory Task 2: Fluorescence Imaging of Focal Adhesion
References
Chapter 8: Contact Guidance and Cell Patterning
8.1 What Is Contact Guidance?
8.2 Contact Guidance on Basement Membrane
8.3 Nanogrooves
8.4 Shear Flow-Resistant Composite Nanosurfaces
8.5 Contact Guidance by Flow
8.6 Cell Patterning via Lithographic Protein Patterns
8.7 Cell Patterning via Direct Protein Deposition
8.8 Cell Patterning via Microcontact Printing
8.9 Direct Cell Patterning via Inkjet Printing (Cell Printing)
8.10 Laboratory Task 1: Contact Guidance on Microgroove
Objective 1. Simple Fabrication of a Microgroove
Objective 2. Seeding Cells on a Microgroove
Objective 3. Fluorescence Imaging of Cells on a Microgroove
8.11 Laboratory Task 2: Cell Patterning via Droplet Collagen Deposition
References
Chapter 9: 3D Scaffold Fabrication
9.1 3D Scaffolds
9.2 Electrospinning
9.3 Materials Selection for Electrospinning Toward Tissue Engineering
9.4 Modification of Electrospun Fibers for Tissue Engineering
9.5 Contact Guidance on Electrospun Fibers
9.6 Controlled Release of Growth Factors and Bioactive Factors from Electrospun Fibers
9.7 Core–Shell Structured Electrospun Fibers
9.8 Addition of GAG-Like Structure to Electrospun Fibers
9.9 3D Printed Scaffolds
9.10 Various 3D Printing Methods
9.11 Hydrogel Bioprinting
9.12 Adding Porosity and Nanostructure to 3D Printing
9.13 Indirect 3D Printing
9.14 Laboratory Task 1: Electrospinning
Objective 1. Preparation of Electrospinning
Objective 2. Electrospinning
Objective 3. Characterization of Electrospun Fibers
9.15 Laboratory Task 2: 3D Printing
References
Chapter 10: Design of In Vitro Culture and Bioreactor
10.1 Design of In Vitro Culture
10.2 Doubling Time (td)
10.3 Mean Residence Time (tres)
10.4 Oxygen Depletion Time (tO2dep)
10.5 Oxygen Diffusion Time (tO2diff)
10.6 Design of In Vitro Culture Considering Both Oxygen Depletion Time and Oxygen Diffusion Time
10.7 Design of Tissue-Engineered Device Using Characteristic Time
10.8 Tissue Engineering Bioreactor
10.9 Measurements from a Tissue Engineering Bioreactor
10.10 Laboratory Task 1: Design of In Vitro Culture and Co-culture
10.11 Laboratory Task 2: Design of Artificial Vasculature Using Oxygen Depletion and Diffusion Times
References
Chapter 11: Organ-on-a-Chip
11.1 2D Versus 3D Cell Culture
11.2 Organ-on-a-Chip
11.3 How Do You Fabricate Lab-on-a-Chip (LOC)?
11.4 OOC Example: Kidney-on-a-Chip
11.5 OOC Example: Liver-on-a-Chip
11.6 OOC Example: Lung-on-a-Chip
11.7 OOC Example: Angiogenesis-on-a-Chip
11.8 OOC Example: Blood–Brain Barrier (BBB)-on-a-Chip
11.9 Other OOC Examples
11.10 Multiple-Organs-on-a-Chip and Human-on-a-Chip
11.11 OOC Application: Drug Testing
11.12 OOC Application: Disease Model
11.13 Mechanical Stimuli to OOCs
11.14 Laboratory Task 1: OOC Fabrication
11.15 Laboratory Task 2: Drug Testing with OOC
References
Chapter 12: Tissue-Engineered Skin Transplant
12.1 When Do You Need Skin Transplants?
12.2 Basic Anatomy of Skin
12.3 How Can We Culture Keratinocytes In Vitro?
12.4 Langerhans Cells and Immune Response of Skin
12.5 Scaffold for Tissue-Engineered Skin Transplant
12.6 Use of Stem Cells for TE Skin Transplant
12.7 Laboratory Task 1: Construction of Skin TE Scaffold
12.8 Laboratory Task 2: Seeding and Proliferating Keratinocytes on the TE Skin Scaffold
12.9 Laboratory Task 3: Force: Deflection Curve of the TE Skin Transplant
References
Chapter 13: Vascularization of Tissue Transplants
13.1 Angiogenesis and Vascularization
13.2 Anatomy and Physiology of Blood Vessel
13.3 Process of Angiogenesis
13.4 Angiogenesis-Stimulating Growth Factors and Angiogenesis Inhibitors
13.5 HUVEC
13.6 Formation of Vasculature Network
13.7 Alignment of Vascular Endothelial Cells to Flow
13.8 Laboratory Task 1: Vessel Sprouting by VEGF on Paper-Based Model
Objective 1. HUVEC Culture
Objective 2. Cell Patterning on Paper
Objective 3. Addition of VEGF
Objective 4. Fluorescence Imaging
13.9 Laboratory Task 2 (Optional): Vessel Sprouting by Mechanical Stimuli on Paper Model
References
Chapter 14: Advanced Topics
14.1 Cartilage Tissue Engineering
14.2 Bone Marrow Transplantation
14.3 Cardiac Patches
14.4 Immunoisolated Pancreas
14.5 Kidney Tissue Engineering
14.6 Other TE Transplants
References
Index