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  • Cited by 23
Publisher:
Cambridge University Press
Online publication date:
July 2010
Print publication year:
2010
Online ISBN:
9780511730207

Book description

Protecting the reproductive potential of young patients undergoing cancer therapy is increasingly important. With modern treatment protocols, 80% of patients can be expected to survive. It has been estimated that up to one in 250 young adults will be a survivor of childhood cancer in the future; infertility, however, may be a consequence. As a wide range of fertility preservation methods are increasingly offered by clinicians, this systematic and comprehensive textbook dealing with the cryobiology, technology and clinical approach to this therapy will be essential reading to infertility specialists, embryologists, oncologists, cryobiologists, ObGyns, andrologists, and urologists with an interest in fertility preservation. Fertility Cryopreservation reviews all the techniques of this increasingly important field within reproductive medicine. It covers the basic principles of pertinent cryobiology, and contains major sections on the different therapies available, written by international specialists combining experience from both academic centers and commercial industries.

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Contents


Page 2 of 2


  • 20 - Obstetric and perinatal outcomes in pregnancies conceived following oocyte cryopreservation
    pp 178-188
  • View abstract

    Summary

    Supernumerary embryos may be cryopreserved as early as day 1 (pronuclear stage) and up to day 6 or day 7. This chapter focuses on the crypreservation of pronuclear stage embryos on day 1. Two main methods are used for the cryopreservation of human oocytes and embryos: slow freezing and vitrification. Although controlled slow freezing remains the main method of cryopreservation in most in vitro fertilization (IVF) programs, the vitrification technique has entered the mainstream of human assisted reproductive technology (ART) more and more. Successful pregnancies and live births have been achieved by vitrification of human oocytes and embryos using different carrier systems. The total concentration of cryoprotectant ranges from 5 to 8 mol/l in vitrification protocols currently used in the cryopreservation of human oocyte and embryos. Relatively limited data are available regarding the application of vitrification in the cryopreservation of pronuclear stage human embryos.
  • Section 5 - Cryopreservation of ovarian tissue
    pp 189-247
  • View abstract

    Summary

    Cryopreservation of human embryos is an important tool for in vitro fertilization (IVF) practice. Human embryos can be cryopreserved efficiently from the pronuclear to blastocyst stage. For day 2 or day 3 embryos, it is better practically to culture the thawed embryos further for a period of time before transfer in order to confirm that these embryos resume development. The selection method for post-thawed embryos for transfer directly affects the clinical outcome in terms of pregnancy and live birth rates. Two basic techniques have been employed for the cryopreservation of human embryos: slow freezing and vitrification. Slow freezing was used first and is still the most common method for cryopreservation of human embryos. Vitrification is a relatively new technique and has been introduced to cryopreserve human embryos in recent years. Many studies have reported improved success rates in terms of clinical pregnancy and live birth rates with the vitrification technique.
  • 22 - In vitro culture of human primordial follicles
    pp 200-212
  • View abstract

    Summary

    Cryopreservation techniques for blastocysts are required to preserve surplus blastocysts. Cryopreservation of the blastocyst was mainly carried out with a slow-freezing technique using programmed freezers. As a typical container for embryos, a cryostraw, includes both liquid (cryoprotectant) and solids, a slow change in temperature is needed to preserve cell structure. The vitrification method developed for mammals has recently been applied to human embryos. The chapter outlines a vitrification procedure for cryopreservation of blastocysts (Cryotop method). The cryopreservation of surplus blastocysts is beneficial, especially for day 6 blastocysts. When cryopreserved and thawed blastocysts are transferred into the uterus, two methods of endometrial preparation for transfer are available. One is transferring to the endometrium in the natural menstrual cycle and the other is to an endometrium prepared by hormone replacement. For patients who have a chromosomal abnormality, preimplantation genetic diagnosis (PGD) is applied for in order to choose normal embryos.
  • 23 - Concept of human ovarian tissue cryobanking
    pp 213-217
  • View abstract

    Summary

    This chapter discuses the protocol development and clinical application of aseptic vitrification of human blastocysts. For non-aseptic embryo carrier, the Hemi-straw is used as an embryo carrier device. Vitrisafe is used as an aseptic embryo carrier. The chapter presents the results for the development of an aseptic vitrification protocol. Before an aseptic vitrification is implemented into a routine clinical program, two issues have to be solved to design an embryo carrier device allowing cooling and storage without contact with LN2. The first issue is to be able to achieve and maintain conditions within the embryos that guarantee an amorphous state throughout the cooling as well during the warming process. The second issue concerns the determination of a protocol for exposing blastocysts to cryoprotectants (CPAs) before vitrification in conditions where cooling rates are reduced because of the heat-insulating barrier of the straw in which the blastocysts are kept.
  • 24 - Transplantation of cryopreserved ovarian tissues
    pp 218-232
  • View abstract

    Summary

    An effective oocyte cryopreservation program benefits infertile couples with moral or religious objections about cryopreservation of embryos. When considering all pregnancies and live births obtained from cryopreserved oocytes using the classic slow-freezing method, the survival rates averaged approximately 50%. The percentage of live births per thawed egg ranges from 1 to 10% using the classic slow-freezing protocols. Recently, improved survival and pregnancy rates have been reported using modified slow-freezing procedures, particularly increased sucrose concentration in the suspending solution, and the use of sodium-free freezing solutions. Several attempts have been made with immature human oocytes. Although survival rates seemed to be improved by the slow-freezing method, poor in vitro maturation (IVM) and fertilization are major problems associated with immature egg freezing. Rapid cooling (vitrification) of human oocytes has resulted in relatively higher survival rates. This study suggested that better results can be achieved by vitrifying mature oocytes rather than immature oocytes.
  • 26 - Transplantation of whole frozen–thawed ovaries
    pp 241-247
  • View abstract

    Summary

    Oocyte cryopreservation has long been envisaged as the ideal solution to the need to capitalize the reproductive potential derived from a cycle of ovarian stimulation, circumventing the ethical and legal problems posed by embryo cryopreservation. This chapter presents an overview of the current status of oocyte cryopreservation by slow-cooling methodology. It discusses recent advances, unresolved matters, and possible future developments. Cryoprotection may be achieved through the use of cryoprotective agents (CPAs), chemicals that interfere with the water-ice transition and interact with biomolecules, acting as water replacement. The use of alternative and less toxic CPAs is a potential approach in the development of more efficient slow-cooling protocols. Assessment of oocyte quality is one of the most crucial, intriguing and, yet, unresolved questions in vitro fertilization (IVF) and gamete biology. Ooplasmic vacuolization is frequently detected in mammalian oocytes treated with different cryopreservation protocols or exposed to CPA alone.
  • Ethical considerations
    pp 248-256
  • View abstract

    Summary

    This chapter focuses on the effects of using different types of a physical device, the carrier tool, to carry or enclose the cellular sample in the process of vitrification of human oocytes. It discusses the effectiveness and safety of various tools used in vitrification to hold the oocytes during cooling, storage, and warming. Cooling and warming rates play a critical role in the success of vitrification in assisted reproductive technology (ART) and, in particular, with human oocytes, which appear to be more sensitive to cryopreservation than preimplantation embryonic stages. Cryostorage is the only situation in ART where numerous samples from multiple patients are pooled together in a common liquid environment. Ethylene glycol is a standard part of all present vitrification protocols. Combinations of cryoprotectant (CPAs) have been tested, and finally a mix of ethylene glycol and dimethyl sulfoxide seems to be the most popular choice.
  • Appendix 1 - Autotransplantation of cryopreserved thawed human ovarian tissue
    pp 257-259
  • View abstract

    Summary

    Oocyte cryopreservation represents an attractive and the least invasive option of fertility preservation strategy for patients wishing to retain their choice of a male partner. Recent advances in vitrification techniques have markedly improved the efficacy of oocyte cryopreservation, demonstrating that vitrification may be more effective than the conventional slow-cooling method. The proposed mechanism of vitrification uses a high concentration of cryoprotectant (CPA) and extremely rapid cooling and warming rates in order to avoid intra-and extracellular ice formation. The commonly used CPAs for cryopreservation of oocytes and embryos are small neutral solutes such as ethylene glycol (EG), 1,2-propanediol (PROH), dimethyl sulfoxide (DMSO), and glycerol. Some devices used for vitrification include the McGill cryoleaf, cryotip, cryotop, nylon loop or cryoloop, thin capillary or open pulled straws, hemi-straws, and electron microscope copper/gold grids. The oocyte vitrification technique offers a realistic hope of fertility preservation for women without partners.

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