1st Edition
3D Cell Culture Fundamentals and Applications in Tissue Engineering and Regenerative Medicine
3D cell culture is yet to be adopted and exploited to its full potential. It promises to upgrade and bring our understanding about human physiology to the highest level with the scope of applying the knowledge for better diagnosis as well as therapeutics. The focus of this book is on the direct impact of novel technologies and their evolution into viable products for the benefit of human race. It also describes the fundamentals of cell microenvironment to bring forth the relevance of 3D cell culture in tissue engineering and regenerative medicine. It discusses the extracellular matrix/microenvironment (ECM) and emphasizes its significance for growing cells in 3D to accomplish physiologically viable cell mass/tissue ex vivo. The book bridges the knowledge gaps between medical need and the technological applications through illustrations. It discusses the available models for 3D cell culture as well as the techniques to create substrates and scaffolds for achieving desired 3D microenvironment.
Preface
1. Introduction
1.1. Cell Culture: Historic Perspective
1.2. Cell Culture; 2D vs. 3D
References
2. Significance of Extracellular Matrix/Microenvironment (ECM)
2.1. Introduction
2.1.1. What is ECM?
2.1.2. Tissue Specificity of ECM
2.1.3. Representative Components of ECM
2.1.3.1. Collagen (structural)
2.1.3.2. Elastin (stretchable)
2.1.3.3. Microfibril-associated macromolecules (fibrillar)
(i) Microfibril-associated glycoproteins (MAGPs)
(ii) Fibrillin
(iii) Fibulins
(iv) Elastin–microfibril interface located proteins (EMILIN)
2.1.3.4. Laminin (adhesive)
2.1.3.5. Fibronectin (adhesive)
2.1.3.6. Matricelluar (antiadhesive)
2.1.3.7. Matrikines and Matricryptins
2.1.3.8. Proteoglycans
(i) Small Leucine-rich Proteoglycans (SLRP)
(ii) Modular Proteoglycans
(a) Nonhyaluronan binding
(b) Hyalectans
(iii) Cell-surface/transmembrane proteoglycans
(a) Syndecans
(b) Glypicans
2.1.3.9. Glycosaminoglycans (GAGs)
2.1.3.10. Hyaluronan (Hyaluronic acid)
2.2. Cell–Cell Interaction
2.3. Cell–Effecter Interaction
2.4. Cell–ECM Interaction
2.4.1. Protrusive Contacts
2.4.2. Contractile Contacts
2.4.3. Mechanically Supportive Contacts
2.5. ECM-Related Disorders
2.6. Conclusion (Classification, Interactions, and Implications)
References
3. ECM-Mimicking for 3D Cell Culture
3.1. Introduction
3.1.1. Significance of ECM Mimicking
3.1.2. ECM Mimicking/Reconstitution
3.1.2.1. Physical Shape and Morphology
3.1.2.2. Biochemical Attributes
3.1.2.3. Mechanical Strength and Elasticity
3.1.2.4. Organ Decellularization
3.1.3. Compatible Cell Types
3.2. Models for 3D Cell Culture
3.2.1. Spheroids
3.2.2. Hydrogels
3.2.3. Scaffolds and Matrices
3.3. Materials for 3D Cell Culture
3.3.1. Natural
3.3.2. Synthetic (Inorganic and Organic)
3.3.3. Hybrid
3.3.3.1. Physical blends
3.3.3.2. Chemical composites
3.4. Methods for Creating 3D Scaffold/Matrices
3.4.1. Conventional Methods
3.4.1.1. Extrusion
3.4.1.2. Compression molding with particle leaching
3.4.1.3. Injection molding
3.4.1.4. Thermally-induced phase transition/gas foaming
3.4.1.5. Fiber bonding and Microsphere sintering
3.4.1.6. Gel casting and freeze drying
3.4.1.7. Solvent casting/melt molding and particulate leaching
3.4.1.8. Micro-contact printing
3.4.2. Advanced Methods
3.4.2.1. Electrospinning
3.4.2.2. Rapid prototyping or solid free form fabrication
(i) Stereolithography/3D laser lithography
(ii) CAD-based 3D plotting
(iii) CAD-based 3D printing
(iv) 3D Fiber/Fused deposition
(v) Selective laser sintering(SLS)
3.4.2.3. Emulsion templating
3.4.2.4. Micromolding
3.4.2.5. Photoplating/photolithography
3.4.2.6. Designed self-assembly
3.5. Applications of ECM-mimicking 3D scaffolds
3.5.1. Research and Development
3.5.2. Diagnostics
3.5.3. Cell-Based Sensors
3.5.4. High-Throughput Screening
3.5.5. Biotech Industry
3.5.6. Drug Delivery
3.5.7. Biochemical Replacement
3.5.8. Tissue Engineering
3.5.8.1. Ex vivo Organ Model
3.5.8.2. Tissue Explants
3.5.8.3. In vivo Tissue Regeneration
3.5.9. Human-Organoid Models
References
4. Types of Scaffolds in Cell/Tissue Culture
4.1. Introduction
4.2. Non-specific in vitro 3D Culture
4.3. Tissue Engineering
4.4. Regenerative Medicine
4.4.1. In situ
4.4.2. Ex situ
4.5. Available Technologies
4.5.1. Matrigel
4.5.2. Alvetex (Re-innervate)
4.5.3. 3D Biotek
4.5.4. Algimatrix
4.5.5. Puramatrix
4.5.6. Integra
4.5.7. Primatrix
4.5.8. Hyalubrix/Hyalgan
4.5.9. Extracel
4.5.10. Mebiol
4.5.11. UpCell
4.5.12. BioVaSc-TERM®
4.5.13. Corgel TrelX/TrelXC
4.5.14. Opsite, Biobrane, and Oasis® Wound Matrix
4.5.15. Cytomatrix
4.5.16. Amniograft
4.5.17. Artiss/Tisseel
4.5.18. ECM analog
References
5. Future Trends/Challenges Ahead
6. Acknowledgement
7. Recommended Reading
8. Glossary
Biography
Ranjna C. Dutta is the founder director of ExCel Matrix Biological Devices Pvt. Ltd., Hyderabad, India, and manager at Centre of Excellence—Biomaterials for Orthopedic and Dental Applications, Material Research Centre, Indian Institute of Sciences (IISc), Bangalore, India. She was awarded a PhD by Central Drug Research Institute, Lucknow, India. During her PhD days, she started working on the synthesis and evaluation of immune-modulating peptides for targeting liposomes to macrophages as a biomedical researcher and ended up joining hands with Dr. Aroop K. Dutta, as founder director of India’s first and only innovation-based start-up in tissue engineering and regenerative medicine. Post PhD she worked at the National Institute of Immunology and the International Center for Genetic Engineering and Biotechnology in New Delhi, India, and the Indian Institute of Chemical Technology (IICT) and the National Institute of Nutrition in Hyderabad, India, on various collaborative as well as her own projects. She has been a visiting post-doctoral fellow in the Northwestern University, Illinois, USA. One of her innovative projects got her Prolog to Discovery award from IICT. For one of her patents, coauthored with Dr. A. K. Dutta, earned her a gold medal from DST-Lockheed Martin Innovation Growth Program–2009 and the Leaders in Innovation Fellowship–2015 from Royal Academy of Engineering, UK, and which is also being actively being pursued for novel product development. Dr. Dutta has over 22 International publications with more than 350 citations. For the last couple of years she has been engaged in the design and synthesis of novel, well-defined ECM mimicking macro-conjugates that can be used for creating instructive 3D scaffolds.
Aroop K. Dutta is the managing director of ExCel Matrix Biological Devices Pvt. Ltd. He is a biochemical engineer by profession and received his B.Tech degree from Harcourt Butler Technological Institute, Kanpur, India, and his M.Tech degree and PhD from the Indian Institute of Technology, Kharagpur, India. After working with both small and large pharmaceutical players, Dr. Dutta was convinced of his unsuitability to the generic environment and he decided to follow his heart to pursue innovative R&D as regenerative medicine start-up. He did bet on his original research and postdoctoral work and obtained exclusive license of the technology that he had helped to develop in his parent institute. ExCel Matrix was the consequential venture. His vision is to develop innovative 3D cell culture technologies for regenerative medicine and conceptualize relevant business models to commercialize such technologies in India. Currently, he is involved with application development of ECM Analog® and Sol-Cell-Gel® technologies and their commercialization in India. Academically, Dr. Dutta is passionate about interfacing biology and engineering for healthcare applications. He has pioneered innovative rapid tissue prototypes and tissue manufacturing concepts and his key strategic focus has been on socio-economic challenges to innovations in a developing economy. His other passion is pedagogy and equity participation by the society around innovations. His greatest dream is to build a "Skunk Works" team and develop bioengineering and regenerative medicine products.
