Chapter
Mammalian Cells: Modern Workhorses
Cells as Therapeutic Agents
Biomarkers for Health or Disease
One Size Does Not Fit All
The Process is the Product
Dispelling the Myth of the Cold Black Box
Chapter 2 Pre-freeze Processing and Characterization
Digestion of Cells from Intact Tissue
Selection of Subpopulations
Activation or Stimulation
Pre-freeze Process Monitoring
Pre-freeze Characterization
Microbial Testing of Cell Therapy Products
Special Considerations for the Characterization of Cell Therapies
Annotation of Pre-freeze Processing
Putting Principles into Action
Chapter 3 Formulation and Introduction of Cryopreservation Solutions
Importance of Cryoprotective Agents
Mechanisms of Cryoprotection
Formulating a Cryopreservation Solution
Formulation of a Vitrification Solution
Characterization and Quality Control for Cryoprotective Solutions
Developing a Protocol for Introducing CPA Solutions
Introduction of Vitrification Solutions
Safety Considerations for Cryopreservation Solutions
Cryopreservation Containers
Putting Principles into Practice
Chapter 4 Freezing Protocols
Importance of Cooling Rate
Controlled Cooling-rate Protocols
Segment 1: Initial Hold Period
Verifying Segment 2 (Including S2a)
Other Types of Controlled-rate Protocols
Independent Temperature Measurement
Putting Principles into Practice
Chapter 5 Storage and Shipping of Frozen Cells
Scientific Basis for Selection of a Storage Temperature
Additional Considerations for Vitrified Samples
Standards, Guidelines, and Best Practices
Storage Equipment and Environment
Mapping Storage Devices and Setting Alarm Limits
Inventory Management System
Influence of Background Ionizing Radiation on Stability in Storage
Shelf-Life of Samples in Storage
Fit-for-Purpose Storage Practices
Risk Mitigation in Long-Term Storage
Shipping or Transport of Cells
General Shipping Considerations
Liquid Nitrogen Dry Shippers
Temperature Mapping of a Shipper
Packaging of Samples Being Shipped
Putting Principles into Practice
Chapter 6 Thawing and Post-Thaw Processing
Transporting Samples Prior to Thawing
Estimating Your Thawing Rate
Thawing and Infusion of Cell Therapy Products
Safety Considerations for Thawing
Infusion of Cells Immediately Post-Thaw
Removal of Vitrification Solutions
Putting Principles into Practice
Chapter 7 Post-Thaw Assessment
Common Measures Used in Post‐Thaw Assessment
Differentiation Potential
Transplantation Potential
Strategies to Improve the Accuracy and Reproducibility of Post-Thaw Assessment
Eliminate Measurement Bias
Compensating for Post-Thaw Apoptosis
Post-Thaw Assessment Using a Single Measure
Optical Methods of Post‐Thaw Assessment
Putting Principles into Practice
Chapter 8 Algorithm-Driven Protocol Optimization
Small Cell Number/High Throughput Approach
Validating Operation of the Algorithm
Cryopreservation of Endothelial Cells in Suspension
Preparation of Cryoprotectant Solution
Using Pentastarch Solution
Controlled-rate Freezing with a Methanol Bath
Alternative Freezing Procedure
Cryopreservation of Peripheral Blood Mononuclear Cells from Whole Blood
Protocol 1: Isolation of PBMCS Directly over Ficoll-Hypaque
Protocol 2: Isolation of PBMCS Using SepMates
Appendix A Human Serum AB Freezing Media
Cryopreservation of Human Adipose Stem Cells
Isolation of Human ASCs from Lipoaspirate
Magnetic Cell Sorting (Optional)
Controlled-rate Freezing of Human ASCs
Cryopreservation of Red Blood Cells
MethodI: High Glycerol/Slow Cooling Technique (Meryman and Hornblower 1972)
Preparation of the RBC Concentrate
Addition of the Cryoprotective Solution
Removal of the Cryoprotectant and Debris
MethodII: A Low Glycerol/Rapid Cooling Technique (Rowe, Eyster, and Kellner 1968)
MethodIII: Hydroxyethylstarch/Rapid Cooling Technique (Sputtek 2007)
Cryopreservation of Oocytes by Slow Freezing
Equipmentand Supplies Needed
Oocyte Vitrification and Warming
Transportation of Hematopoietic Progenitor Cells and Other Cellular Products
Cryopreservation of Hematopoietic Progenitor Cells
Appendix A Alternate Cryopreservation HarnessSet-2 or 4 Bags
Thawing of Hematopoietic Progenitor Cells
Processing and Cryopreservation of T-Cells
Protocol/Processing Schema: N/A
Thawing and Reinfusion of Cryopreserved T-Cells
Protocol/Processing Schema
Preservation of Cells
：A Practical Manual
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