Chapter 11
Utilization of the Mixed Lymphocyte Reaction (MLR) Assay to Determine Stem Cell Immunogenicity and Suppression

11.1 Introduction 11.2 Experimental Design 11.2.1 Immunogenicity Assay 11.2.2 Suppression Assay 11.2.3 T Cell Priming Assay 11.3 Materials 11.4 Methods 11.4.1 General Considerations 11.4.2 Safety 11.4.3 Media Preparation 11.4.4 Prepare Responder Cell Populations 11.4.5 Prepare Stimulator Cell Populations 11.4.6 Performance of Immunogenicity Assays 11.4.7 Performance of Suppression Assays 11.4.8 Performance of T Cell Priming Assays 11.4.9 MLR plate culture, pulsing with 3H-thymidine, cell harvest and scintillation counting Applicable to all three MLR assays 11.5 Data Acquisition, Anticipated Results, Interpretation and Statistical guidelines 11.5.1 Immunogenicity Assay 11.5.2 Suppression Assay 11.5.3 T Cell Priming Assay 11.6 Discussion and Commentary 11.7 Troubleshooting Table 11.8 Application Notes 11.9 Summary Points

Chapter 12
A Novel Method for the Preservation of Embryonic Stem Cells Using a Quartz Capillary Freezing System

12.1		Introduction
12.1.1		Slow Freezing Protocols
12.1.2		Vitrification Protocols
12.1.3		Increasing Cooling Rates for Vitrification
12.1.4		Quartz Capillary System
12.1.5		Apparent vitrification of CPA-laden solutions
12.1.6		Slush Nitrogen
12.1.7		Experimental Design
12.2		Materials
12.3		Methods
12.3.1		Murine embryonic stem (ES) cell culture
12.3.2		Preparing Slush Nitrogen
12.3.3		Cryopreservation of Murine Embryonic Stem Cells by vitrification
12.4		Anticipated results
12.4.1		Cell attachment and Proliferation Post Vitrification
12.4.2		Pluripotent properties of ES cells Post Vitrification
12.5		Discussion and Commentary
12.6		Troubleshooting Table
12.7		Application Notes
12.8		Summary Points

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