Details

Preservation of Cells


Preservation of Cells

A Practical Manual
1. Aufl.

von: Allison Hubel

76,99 €

Verlag: Wiley-Blackwell
Format: PDF
Veröffentl.: 04.12.2017
ISBN/EAN: 9781118989852
Sprache: englisch
Anzahl Seiten: 216

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Beschreibungen

Helps those that use cell preservation to develop new protocols or improve existing protocols  This book provides readers with the tools needed to develop or debug a preservation protocol for cells. The core structure and content of the text grew from a professional short course that has been offered at the Biopreservation Core Resource for the last 10 years. This comprehensive text describes, step by step, the individual elements of a protocol, including the relevant scientific principles for each phase of the protocol. It can be used by anyone who is involved in cell preservation—even by those who are not experts in freezing of cells—because it provides the scientific basis for those that want to understand the basis for the protocol. Preservation of Cells: A Practical Manual begins by first introducing readers to the subject of preserving cells. It then goes on to cover Pre-freeze Processing and Characterization; Formulation and Introduction of Cryopreservation Solutions; Freezing Protocols; Storage and Shipping of Frozen Cells; Thawing and Post Thaw Processing; Post-thaw Assessment; and Algorithm-driven Protocol Optimization. Clearly explains the reasons behind every step in the development of a preservation protocol and the scientific principles behind them Provides alternative modes of preservation for when conventional methods of cryopreservation are not appropriate for a given cell type or application Enables more organization to achieve improved post thaw recoveries and process consistency  Preservation of Cells: A Practical Manual is an important book for researchers, laboratory technicians and students in cell biology, stem cell biology, tissue engineering, and regenerative medicine. It is also useful to cell bankers, regenerative medicine, biomarker discovery or precision medicine companies, and cell therapy labs, blood bankers, biobankers, and biotechnology companies.
Preface xiii Acknowledgments xvii Nomenclature xix 1 Introduction 1 Mammalian Cells: Modern Workhorses 1 Products from Cells 1 Cells as Therapeutic Agents 2 Biomarkers for Health or Disease 2 In Vitro Models 3 Bridging the Gap 3 The Preservation Toolkit 5 Hypothermic Storage 5 Cryopreservation 6 Vitrification 7 Dry State Storage 8 Fit?]for?]Purpose 8 One Size Does Not Fit All 9 The Process is the Product 9 Reproducibility 12 Safety 12 Dispelling the Myth of the Cold Black Box 12 References 13 2 Pre?]freeze Processing and Characterization 15 Pre?]freeze Processing 15 Digestion of Cells from Intact Tissue 15 Hypothermic Storage 16 Selection of Subpopulations 17 Activation or Stimulation 18 Genetic Modification 18 Culture 19 Pre?]freeze Process Monitoring 19 Pre?]freeze Characterization 20 Identity 20 Genetic Stability 21 Enumeration 21 Purity 22 Adventitious Agents 22 Microbial Testing of Cell Therapy Products 23 Special Considerations for the Characterization of Cell Therapies 24 Annotation of Pre?]freeze Processing 24 Scientific Principles 25 Putting Principles into Action 25 References 26 3 Formulation and Introduction of Cryopreservation Solutions 29 Importance of Cryoprotective Agents 29 Mechanisms of Cryoprotection 31 Formulating a Cryopreservation Solution 31 Formulation of a Vitrification Solution 33 Characterization and Quality Control for Cryoprotective Solutions 34 Toxicity of CPAs 35 Osmotic Toxicity 35 Biochemical Toxicity 36 Developing a Protocol for Introducing CPA Solutions 37 The Basic Experiment 37 Introduction of Vitrification Solutions 38 Cell Concentration 39 Removal of CPA Solution 40 Safety Considerations for Cryopreservation Solutions 40 Cryopreservation Containers 41 Overwraps 42 Labeling 43 Sample Annotation 44 Scientific Principles 44 Putting Principles into Practice 44 References 44 4 Freezing Protocols 47 Importance of Cooling Rate 47 Controlled?]rate Freezing 48 Controlled Cooling?]rate Protocols 49 Segment 1: Initial Hold Period 49 Segment 2: Cooling 50 Uncontrolled Nucleation 53 Manual Nucleation 54 Automatic Nucleation 54 Verifying Segment 2 (Including S2a) 55 “Delayed” Latent Heat 55 Segment 3 56 Verifying Segment 3 56 Other Types of Controlled?]rate Protocols 57 Passive Freezing 57 Transfer to Storage 59 Vitrification 60 Independent Temperature Measurement 60 Scientific Principles 61 Putting Principles into Practice 62 References 62 5 Storage and Shipping of Frozen Cells 65 Scientific Basis for Selection of a Storage Temperature 65 Additional Considerations for Vitrified Samples 67 Standards, Guidelines, and Best Practices 67 Facilities 68 Storage Equipment and Environment 69 Mapping Storage Devices and Setting Alarm Limits 70 Monitoring Systems 71 Safety 71 Inventory Management System 72 Stability in Storage 72 Temperature Fluctuations 73 Influence of Background Ionizing Radiation on Stability in Storage 74 Shelf?]Life of Samples in Storage 75 Fit?]for?]Purpose Storage Practices 75 Risk Mitigation in Long?]Term Storage 76 Shipping or Transport of Cells 76 General Shipping Considerations 77 Liquid Nitrogen Dry Shippers 78 Temperature Mapping of a Shipper 79 Packaging of Samples Being Shipped 79 Monitoring of Shipments 79 Responsibilities 79 Sample Annotation 80 Scientific Principles 80 Putting Principles into Practice 81 References 81 6 Thawing and Post?]Thaw Processing 85 Thawing Equipment 86 Transporting Samples Prior to Thawing 87 Estimating Your Thawing Rate 87 Thawing and Infusion of Cell Therapy Products 89 Safety Considerations for Thawing 90 Post?]Thaw Processing 90 Post?]Thaw Washing 90 Dilution 91 Infusion of Cells Immediately Post?]Thaw 91 Removal of Vitrification Solutions 92 Wash Solutions 92 Scientific Principles 94 Putting Principles into Practice 94 References 94 7 Post?]Thaw Assessment 97 Common Measures Used in Post?]Thaw Assessment 98 Physical Integrity 98 Metabolic Activity 99 Mechanical Activity 100 Mitotic Activity 101 Differentiation Potential 102 Transplantation Potential 103 Strategies to Improve the Accuracy and Reproducibility of Post?]Thaw Assessment 103 Eliminate Measurement Bias 103 Compensating for Post?]Thaw Apoptosis 105 Post?]Thaw Assessment Using a Single Measure 106 Optical Methods of Post?]Thaw Assessment 106 Release Criteria 107 Scientific Principles 107 Putting Principles into Practice 107 References 108 8 Algorithm?]Driven Protocol Optimization 111 Small Cell Number/High Throughput Approach 113 Validating Operation of the Algorithm 114 Flexibility 115 Practical Notes 115 Modeling in Cryobiology 115 References 116 Protocols Introduction 117 Protocol Contributors 118 Cryopreservation of Endothelial Cells in Suspension 119 Principle 119 Equipment and Supplies 119 Equipment 119 Supplies 120 Safety 120 Procedure 121 Cell Preparation 121 Preparation of Cryoprotectant Solution 121 Using Powdered HES 122 Using Pentastarch Solution 122 Cryoprotectant Addition 122 Freezing 122 Controlled?]rate Freezing with a Methanol Bath 122 Alternative Freezing Procedure 123 Thawing 123 Expected Results 123 References 123 Cryopreservation of Peripheral Blood Mononuclear Cells from Whole Blood 125 Principle 125 Protocol 1: Isolation of PBMCS Directly over Ficoll?]Hypaque 125 Equipment 125 Materials 126 Reagents 126 Procedure 126 Protocol 2: Isolation of PBMCS Using SepMates 127 Equipment 127 Materials 128 Reagents 128 Procedure 128 Appendix A Human Serum AB Freezing Media 129 Materials 129 Equipment 130 Reagents 130 Procedure 130 Cryopreservation of Human Adipose Stem Cells 131 Principle 131 Equipment and Supplies 131 Reagents and Media 132 Procedure 133 Isolation of Human ASCs from Lipoaspirate 133 Magnetic Cell Sorting (Optional) 135 Cryopreservation 135 Controlled?]rate Freezing of Human ASCs 135 Thawing Human ASCs 137 Notes 138 Reference 139 Cryopreservation of Red Blood Cells 141 Method I: High Glycerol/Slow Cooling Technique (Meryman and Hornblower 1972) 141 Preparation of the RBC Concentrate 141 Addition of the Cryoprotective Solution 141 Cooling 142 Rewarming 142 Removal of the Cryoprotectant and Debris 142 Method II: A Low Glycerol/Rapid Cooling Technique (Rowe, Eyster, and Kellner 1968) 143 Method III: Hydroxyethylstarch/Rapid Cooling Technique (Sputtek 2007) 144 References 146 Cryopreservation of Oocytes by Slow Freezing 147 Principle 147 Specimen Requirements 147 Equipment and Supplies Needed 147 Equipment 147 Supplies 148 Procedure 148 Safety 152 Calculations 152 Reporting Results 152 Procedure Notes 153 Limitations of Procedure 153 Oocyte Vitrification and Warming 155 Principle 155 Equipment and Supplies 155 Equipment 155 Supplies 155 Procedure 156 Quality Control 160 Safety 161 Transportation of Hematopoietic Progenitor Cells and Other Cellular Products 163 Principle/Rationale 163 Specimen 163 Equipment/Reagents 163 Quality Control 164 Procedure 164 Additional Information 165 Further Reading 165 Cryopreservation of Hematopoietic Progenitor Cells 167 Principle/Rationale 167 Protocol/Processing Schema 168 Specimen 168 Equipment/Reagents 168 QualityControl 169 Procedure169 Appendix A Alternate Cryopreservation Harness Set?]2 or 4 Bags 173 Further Reading 173 Thawing of Hematopoietic Progenitor Cells 175 Principle/Rationale 175 Equipment/Reagents 175 QualityControl 176 Procedure176 Further Reading 177 Processing and Cryopreservation of T?]Cells 179 Principle/Rationale 179 Protocol/Processing Schema: N/A 179 Specimens179 Equipment/Reagents 179 Quality Control 180 Procedure 180 Further Reading 182 Thawing and Reinfusion of Cryopreserved T?]Cells 183 Principle/Rationale 183 Protocol/Processing Schema 183 Specimen 184 Equipment/Reagents 184 Quality Control 184 Procedure 184 Further Reading 187 Index 189
ALLISON HUBEL is a Professor of Mechanical Engineering and the Director of BioCoR (Biopreservation Core Resource) at the University of Minnesota, USA.
Helps those that use cell preservation to develop new protocols or improve existing protocols This book provides readers with the tools needed to develop or debug a preservation protocol for cells. The core structure and content of the text grew from a professional short course that has been offered at the Biopreservation Core Resource for the last 10 years. This comprehensive text describes, step by step, the individual elements of a protocol, including the relevant scientific principles for each phase of the protocol. It can be used by anyone who is involved in cell preservation—even by those who are not experts in freezing of cells—because it provides the scientific basis for those that want to understand the basis for the protocol. Preservation of Cells: A Practical Manual begins by first introducing readers to the subject of preserving cells. It then goes on to cover Pre-freeze Processing and Characterization; Formulation and Introduction of Cryopreservation Solutions; Freezing Protocols; Storage and Shipping of Frozen Cells; Thawing and Post Thaw Processing; Post-thaw Assessment; and Algorithm-driven Protocol Optimization. Clearly explains the reasons behind every step in the development of a preservation protocol and the scientific principles behind them Provides alternative modes of preservation for when conventional methods of cryopreservation are not appropriate for a given cell type or application Enables more organization to achieve improved post thaw recoveries and process consistency Preservation of Cells: A Practical Manual is an important book for researchers, laboratory technicians and students in cell biology, stem cell biology, tissue engineering, and regenerative medicine. It is also useful to cell bankers, regenerative medicine, biomarker discovery or precision medicine companies, and cell therapy labs, blood bankers, biobankers, and biotechnology companies.

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