Chapter 9
Micromolded Nonadhesive Hydrogels for the Self-Assembly of Scaffold-Free 3D Cellular Microtissues

9.1 Introduction 9.2 Experimental Design 9.3 Materials 9.4 Methods 9.4.1 Design and fabrication of micromolds 9.4.2 Casting of agarose micromolded hydrogels 9.4.3 Casting polyacrylamide micromolded hydrogels 9.4.4 Formation of 3D microtissues 9.4.5 Side-on viewing of self-assembly 9.4.6 Spheroid size quantification 9.4.7 Live cell fluorescent staining and determination of self-sorting patterns of cells 9.4.8 WST-1 cell proliferation assay adapted for microtissues 9.4.9 Harvesting microtissues 9.4.10 LIVE/DEAD staining to determine microtissue viability 9.4.11 Data analysis 9.5 Anticipated Results 9.6 Discussion and Commentary 9.7 Application Notes 9.8 Summary Points, Acknowledgments, References

Chapter 10
On-Demand 3D Freeform Fabrication of Tissue Structures Using Bioprinting

10.1		Introduction
10.2		Experimental Design
10.3		Materials
10.3.1		Reagents
10.3.2		Facilities/equipment
10.3.3		Overview of the robotic 3D bioprinter
10.3.4		Software for the 3D bioprinter operations
10.3.5		Sterilization of the fluidic pathways in the 3D bioprinter for biomaterial printing
10.3.6		Loading of materials to the printer
10.3.7		Control of droplet dispensing
10.3.8		Cleaning of the printing path and microvalve after use
10.4		Methods
10.4.1		Optimization of the printing resolution for a cell-laden single-layer hydrogel scaffold
10.4.2		Multilayer cell-laden hydrogel scaffold
10.4.3		Cell-laden hydrogel scaffold on a nonplanar surface
10.4.4		Staining and imaging
10.5		Anticipated Results, Data Acquisition, and Interpretation
10.6		Discussion and Commentary
10.7		Application Notes
10.8		Summary Points, Acknowledgments, References

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