Meet the editor

Marian D. Quain is a biotechnologist whose research focuses on utilization of tissue culture techniques for production of clean planting materials, germplasm conservation and use of molecular tools for crop diversity, disease diagnostics and molecular marker assisted selection breeding using transcriptomics and proteomics, as well as application of recombinant gene technology for crop improvement. Marian was a Visiting Research Fellow

at Leeds University, UK, from 2011 to 2013. In 2013, she was adjudged the National Best Research Scientist by the Ghanaian Ministry of Food and Agriculture (MoFA). She is a 2017 Cochran Fellow and a fellow of the Africa Science Leadership Program. In 2018, she won the American Society for Plant Biologists Award for Excellence in Education for her outstanding contributions to plant biology education.

Contents

**Section 1**

*Samuel Buff and Anne Baudot*

*by Noha A. Al-Otaibi*

in Reproductive Medicine

*by Nabil Sayme*

*and Dayong Gao*

**Section 3**

Techniques

**Section 2**

**Preface III**

Recceing Oligomers, Polymers and Other Cryoprotective Agents **1**

**Chapter 1 3**

**Chapter 2 29**

**Chapter 3 49**

Procedures for Cryopreserving Gametes **61**

**Chapter 4 63**

**Chapter 5 75**

Cryobiology Aiding Organ Transplant **117**

**Chapter 6 119**

The Use of Chitooligosaccharides in Cryopreservation: Discussion

of Concept and First Answers from DSC Thermal Analysis *by Hugo Desnos, Pierre Bruyère, Magda Teixeira, Loris Commin, Gérard Louis, Stephane Trombotto, Amani Moussa, Laurent David,* 

*by Mark D. Scott, Nobu Nakane and Elisabeth Maurer-Spurej*

Cryopreservation of Human Spermatozoa: A New Frontier

*by Ali Erdem Öztürk, Mustafa Numan Bucak, Mustafa Bodu, Nuri Başpınar, İlhami Çelik, Zhiquan Shu, Nazan Keskin* 

Current Advancements in Pancreatic Islet Cryopreservation

*by Samuel Rodriguez, David Whaley, Michael Alexander, Mohammad Rezaa Mohammadi and Jonathan R.T. Lakey*

Cryobiology and Cryopreservation of Sperm

Cryoprotection of Platelets by Grafted Polymers

Cryomedia Formula: Cellular Molecular Perspective

## Contents



Preface

Cryopreservation is the storage of biological material at ultra-low temperatures, preferably that of liquid nitrogen, which arrests all metabolic activities. This technique is widely applied to all organisms ranging from microorganisms through plants to animals and human organs. Theoretically, no genetic changes should occur during cryopreservation, thus permitting indefinite cell preservation.

**Cryopreservation** or **cryoconservation** is a process where organelles, cells, tissues, extracellular matrices, organs, or any other biological constructs susceptible to damage caused by unregulated chemical kinetics are preserved by cooling to very low temperatures ranging from −80 to −210°C using programmable freezers, liquid nitrogen, and liquid nitrogen slush and vapour. A successful cryopreservation procedure therefore entails ensuring the normal functionality of the organism. In recent times, basic cryopreservation has had applications in research and clinical,

Typically, freezing or cooling is lethal to most living organisms' cellular,

functionality of tissue as subjected to cryopreservation.

providing discussions, overviews, and reviews of cryo-techniques.

mechanical, and metabolic functionality. Hence the typical procedures involved in cryopreservation include selecting appropriate tissue, conditioning the tissue, determining cooling rate, storing the tissue, thawing the tissue, and subsequently restoring tissue functions. During cryopreservation, several dynamics come into play; these include cell type, size, water content, temperature, and metabolic state. There have been several limitations to the effective application of cryopreservation techniques and storage, although substantial successes have been reported for both animal and plant cells. Cells have to be protected from damage, especially by ice crystals during freezing. Cryoprotectants, also called Cryo Protective Agents (CPAs), are widely used; however, these may be toxic to the cells depending on the type of cell and the extent of exposure. Mode of freezing, thawing, regenerating, and rejuvenating also may render the process successful or otherwise. Viability of cells may be compromised at any step of the process. Naturally, tissues subjected to cryotemperatures contain fluids such as intracellular and extracellular fluids, plasma, interstitial fluid, and transcellular fluid, which make up 10 to 75 percent of tissues. During cooling, these fluids may form ice crystals, which may nucleate to larger crystals and ultimately damage cells. Hence the use of CPAs may render stability and protection to cell membranes, and where proven useful, preserve

The main objective of this book is to bring to bear factors that affect cells during cryopreservation. Divided into four sections, this book contains eight chapters

The first section is on "Recceing Oligomers, Polymers, and Other Cryoprotective Agents (CPAs)." Successful cooling is dependent on the control of extracellular ice formation, protective intracellular dehydration, and the colligative and dehydration properties of the CPA. Hence CPAs ensure a freeze-avoidance mechanism that enables hydrated tissues to survive when exposed to cryogenic temperatures. There are, however, unanswered questions about the precise mechanism of action of CPAs. In their function to protect living cells against damage during cooling,

medical, and agricultural fields.
