3D Cell Culture
Methods and Protocols

Developed for a range of tissues where the culture environment takes into account the spatial organization of the cells therein, 3D cell culture models serve to bridge the gap between in vivo studies at one extreme with that of simple cell monolayers at the other. In 3D Cell Culture: Methods and Protocols, international experts describe a number of basic and applied methodologies taken from a breadth of scientific and engineering disciplines, many of which deal with direct applications of 3D culture models, most notably in the formation of tissues for clinical purpose. Beginning with an overview of the biological and materials scaffold requirements for successfully creating 3D models, the book delves into topics such as general scaffold design and fabrication techniques, models for bone, skin, cartilage, nerve, bladder, and hair follicles, and chapters on bioreactor design, imaging, and stem cells. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective subjects, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, 3D Cell Culture: Methods and Protocols serves as a basic manual for laboratory-based scientists who not only need to have a comprehensive range of techniques contained within a single text but also require techniques described using a standard, convenient format.

Developed for a range of tissues where the culture environment takes into account the spatial organization of the cells therein, 3D cell culture models serve to bridge the gap between in vivo studies at one extreme with that of simple cell monolayers at the other. In 3D Cell Culture: Methods and Protocols, international experts describe a number of basic and applied methodologies taken from a breadth of scientific and engineering disciplines, many of which deal with direct applications of 3D culture models, most notably in the formation of tissues for clinical purpose. Beginning with an overview of the biological and materials scaffold requirements for successfully creating 3D models, the book delves into topics such as general scaffold design and fabrication techniques, models for bone, skin, cartilage, nerve, bladder, and hair follicles, and chapters on bioreactor design, imaging, and stem cells. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective subjects, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, 3D Cell Culture: Methods and Protocols serves as a basic manual for laboratory-based scientists who not only need to have a comprehensive range of techniques contained within a single text but also require techniques described using a standard, convenient format.

1. 3D Cell Culture: A Review of Current Approaches and Techniques
John W. Haycock

2. Scaffolds for Tissue Engineering and 3D Cell Culture
Eleonora Carletti, Antonella Motta, and Claudio Migliaresi

3. Tracking Nanoparticles in Three Dimensional Tissue Engineered Models Using Confocal Laser Scanning Microscopy
Vanessa Hearnden, Sheila MacNeil, and Giuseppe Battaglia

4. Using Immuno-Scanning Electron Microscopy for the Observation of Cell Adhesion Interaction at the Nano- and Microscale in S-phase Cells
Manus J.P. Biggs, R. Geoff Richards, and Matthew J. Dalby

5. 3D Sample Preparation for Orthopedic Tissue Engineering Bioreactors
Sarah H. Cartmell, Sarah Rathbone, Gemma Jones, and L. Araida Hidalgo-Bastida

6. Quantification of mRNA Using Real-Time PCR and Western Blot Analysis of MAPK Events in Chondrocyte/Agarose Constructs
David A. Lee, June Brand, Donald Salter, Oto-Ola Akanji, and Tina T. Chowdhury

7. Genetic Modification of Chondrocytes Using Viral Vectors
Teresa Coughlan, Aileen Crawford, Paul Hatton, and Michael Barker

8. Stem Cell and Neuron Co-Cultures for the Study of Nerve Regeneration
Paul J. Kingham, Cristina Mantovani, and Giorgio Terenghi

9. Production of Tissue Engineered Skin and Oral Mucosa for Clinical and Experimental Use
Sheila MacNeil, Joanna Shepherd, and Louise Smith

10. Three-Dimensional Alignment of Schwann Cells Using Hydrolysable Microfiber Scaffolds: Strategies for Peripheral Nerve Repair
Celia Murray-Dunning, Sally L. McArthur, Tao Sun, Rob McKean, Anthony J. Ryan, and John W. Haycock

11. Encapsulation of Human Articular Chondrocytes into 3D Hydrogel: Phenotype and Genotype Characterization
Rui C. Pereira, Claudio Gentili, Ranieri Cancedda, Helena S. Azevedo, and Rui L. Reis

12. Micro-Structured Materials and Mechanical Cues in 3D Collagen Gels
James B. Phillips and Robert Brown

13. Organotypic and 3D Reconstructed Cultures of the Human Bladder and Urinary Tract
Claire L. Varley and Jennifer Southgate

14. Ex Vivo Organ Culture of Human Hair Follicles: A Model Epithelial - Neuroectodermal - Mesenchymal Interaction System
Desmond J. Tobin

15. Human Endothelial and Osteoblast Co-Cultures on 3D Biomaterials
Ronald E. Unger, Sven Halstenberg, Anne Sartoris, and C. James Kirkpatrick

16. Assessment of Nanomaterials Cytotoxicity and Internalization
Noha M. Zaki and Nicola Tirelli

17. Practical Aspects of OCT Imaging in Tissue Engineering
Stephen J. Matcher

18. Osteogenic Differentiation of Embryonic Stem Cells in 2D and 3D Culture
Lee Buttery, Robert Bielby, Daniel Howard, and Kevin Shakesheff

19. 3D Structuring of Biocompatible and Biodegradable Polymers via Stereolithography
Andrew A. Gill and Frederik Claeyssens

20. Alvetex®: Polystyrene Scaffold Technology for Routine Three Dimensional Cell Culture
Stefan Przyborski