Cryopreservation in Assisted Reproduction

Zsolt Peter Nagy, MD, PhD, Scientific Director, Reproductive Biology Associates - Prelude, Atlanta, GA, USA

Alex C. Varghese, PhD, Scientific Director, Astra Fertility Group, Mississauga, ON, Canada

Ashok Agarwal, PhD, HCLD (ABB), EMB (ACE), Director of Research, Global Andrology Forum, Moreland Hills, and Emeritus Staff, Cleveland Clinic, Cleveland, OH, USA

PART I Introduction.- Cryopreservation in global perspectives: Editors' thoughts.- Historical Background on Gamete and Embryo Cryopreservation.- PART II Principles and Models.- Principles of Cryopreservation.- Animal Models for Human Oocyte, Embryo, and Tissue Cryopreservation.- Considerations in Immature Oocyte Cryopreservation.- Antioxidants and Antifreeze Proteins in Cryopreservation/Vitrification.- Devitrification Thresholds and Warming Rates of Reproductive Cells.- PART III Methodology and Management.- Human Sperm Slow-freezing.- Human Sperm Vitrification.- Human Epididymal and Testicular Sperm Cryopreservation.- Human Oocyte Slow-freezing.- Human Embryo (Zygote, Cleaved, Blastocysts) Slow-freezing.- Human Oocyte Vitrification.- Human Embryo Vitrification  (zygote/cleavage/blastocyst).- Human Ovarian Tissue Slow-freezing.- Human Ovarian Tissue Vitrification.-Vitrification: implementation and training.- Establishing a Donor Oocyte Cryo Bank Network:  USA Experience.- Establishing a Donor Oocyte Cryo Bank Network: European Experience.- Managing a Sperm Bank: European Experience.- Managing a Sperm Bank: USA Experience.- Managing a central cryo depository.- PART IV Advances in Cryotechnology, Research, and Animal Models.- Artificial Intelligence (AI) and oocyte/embryo assessment in cryopreservation cycles.- Vitrification: A Reliable Method for Cryopreservation of Animal Embryos.- Cryopreservation Effect on Genetic/Epigenetic Function: Neonatal Outcomes.- Fertility Preservation and Genome Resource Banking for Rare and Endangered Animal Species.- End of Ice Age and the beginning of Space Age: The Freeze drying of human gametes.- PART V Expected outcomes and Quality Control.- Quality control and KPIs in cryopreservation: Aspiring for the best results.- Handling and assessing of human oocytes and blastocysts after vitrification and warming.- Health of offspring following cryopreservation of oocytes and embryos.- Elective and Onco-fertility preservation outcomes after oocyte vitrification.- PART VI Multiple vitrification-warming and biopsy procedures on human Embryos.- Fertility preservation in pre-pubertal girls.- Fertility Preservation in Transgender Patients.- Fertility Preservation in pre-adolescent and adolescent males: Recent Developments and New Insights.- PART VII Guidelines, Regulations and Best Practices.- Setting up a cryopreservation lab and selecting media, disposables and systems for vitrification.- Cryopreservation and storage of reproductive tissues: Good practice guidelines.- Transportation of Frozen Gametes and Embryos- Good Practice Guidelines.- Good practice guideline of ovarian tissue transportation for optimizing fertility outcomes.- Disposition of cryopreserved specimen.- European regulations for Reproductive Tissue banks.- Ethical issues related to cryopreserved reproductive specimen.- Legal liability landscape and the person/property divide.- Challenges on safe keeping cryo stored specimen in time of armed conflict: Live observations from the war zone.- PART VIII Industrial breakthroughs in cryopreservation and cryo-storage.- At home sperm banking kits.- Single sperm cryopreservation for azoospermia management.- Keeping cryopreserved specimen safe: Thermographic Imaging.- Robotic cryostorage solution.- Maximizing Donor Egg Efficiency - Artificial Intelligence & Genetically Certified Oocytes.- Automating Vitrification.- EmbryoOptions.- LN2 Sterilization to avoid cross contamination.- Lab on a chip - Cryopreservation.- PART IX Appendices - Protocols.- Oocyte slow freezing protocol.- Embryo slow freezing protocol (zygote/cleavage/blastocyst).- Human ovarian tissue slow freezing protocol.- Proper maintenance of dewars/cryo-storage tanks.0 Human Sperm Vitrification.- SOP on maintaining and managing cryopreserved samples and cryo-tanks in an IVF lab.