Chapter 11
Markers for an In Vitro Skin Substitute

11.1 Introduction 11.2 Experimental Design 11.3 Materials 11.3.1 Human tissue-engineered skin substitute reconstructed by the self-assembly approach 11.4 Methods 11.4.1 Preparation of solutions and materials for the in vitro fabrication of human skin substitutes by the self-assembly approach 11.4.2 In vitro fabrication of human skin substitutes by the self-assembly approach 11.4.3 Tissue preservation and sectioning 11.4.4 Preparation of solutions and materials for immunofluorescence 11.4.5 Immunofluorescent labeling of human skin substitutes 11.4.6 Histological analysis 11.4.7 Transmission electron microscopy 11.4.8 Statistical analysis 11.5 Anticipated Results 11.6 Discussion and Commentary 11.7 Application Notes 11.8 Summary Points, Acknowledgements, References

Chapter 12
3D Culture of Primary Chondrocytes, Cartilage, and Bone/Cartilage Explants in Simulated Microgravity

12.1		Introduction
12.2		Experimental Design
12.2.1		Culture models
12.2.2		The RCCS bioreactor and its operational conditions
12.2.3		Animals
12.3		Materials
12.3.1		Equipment for cell/tissue culture and preparation of samples
12.3.2		Chemicals
12.4		Methods
12.4.1		Preparation of tissue explants
12.4.2		Isolation of chondrocytes
12.4.3		2D culture of isolated chondrocytes (traditional monolayer in static fluid conditions)
12.4.4		3D culture of isolated chondrocytes (homotypic aggregates)
12.4.5		3D culture of fragments of articular cartilage explants
12.4.6		3D culture of undissected, complete proximal tibial epiphyses
12.4.7		Histomorphological study of chondrocytes and cartilage tissue
12.5		Anticipated Results
12.6		Discussion
12.6.1		Discussion of pitfalls
12.6.2		General discussion and commentary
12.7		Application Notes
12.8		Summary Points, Acknowledgments, References

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