Allison Hubel

Preservation of Cells

A Practical Manual

• Gebundenes Buch

Produktbeschreibung

Praktikerbuch für alle, die mittels Zellkonservierung neue Protokolle entwickeln oder bestehende Protokolle verbessern.

Dieses Buch vermittelt das notwendige Rüstzeug, um zellbezogene Konservierungsprotokolle zu entwickeln und fehlerhafte Protokolle zu korrigieren. Die Kernstruktur und -inhalte stammen aus einem kurzen Fachkurs, der in den letzten zehn Jahren am Biopreservation Core Resource angeboten wurde. Das umfassende Praktikerbuch beschreibt Schritt für Schritt die einzelnen Teile eines Protokolls, darunter die relevanten wissenschaftlichen Prinzipien für jede Protokollphase, und richtet sich an alle, die sich mit der Zellkonservierung beschäftigen. Es eignet sich auch für Nichtexperten im Bereich Zell-Freezing, denn es stellt die wissenschaftlichen Grundlagen für das Verständnis von Protokollen vor.

Produktdetails

• Verlag: Wiley & Sons / Wiley-Blackwell
• Artikelnr. des Verlages: 1W118989840
• 1. Auflage
• Seitenzahl: 216
• Erscheinungstermin: 13. Februar 2018
• Englisch
• Abmessung: 231mm x 155mm x 18mm
• Gewicht: 408g
• ISBN-13: 9781118989845
• ISBN-10: 1118989848
• Artikelnr.: 48807432

Autorenporträt

ALLISON HUBEL is a Professor of Mechanical Engineering and the Director of BioCoR (Biopreservation Core Resource) at the University of Minnesota, USA.

Inhaltsangabe

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 Syst