The Art of Vitrification and Thawing

ABSTRACT

Vitrification is a method of cryopreservation (freezing) that is widely used in

assisted reproduction to preserve embryos, oocytes, and sperm. This chapter
provides an overview of the principles and techniques of vitrification, including
the use of cryoprotectants, the freezing process, and the storage and thawing
of vitrified specimens. The chapter also discusses the advantages and
limitations of vitrification compared to other cryopreservation methods, and the
clinical applications of vitrification in assisted reproduction, such as in in vitro
fertilization (IVF), intracytoplasmic sperm injection (ICSI), and egg and sperm
banking. Additionally, this chapter will discuss the recent advances in
vitrification technology and future directions for research in this field. Overall,
this chapter is aimed to provide a comprehensive understanding of the
vitrification process and its practical applications in assisted reproduction.

INTRODUCTION

Cryopreservation is a technique used in assisted reproduction to preserve sperm, eggs, and

embryos at very low temperatures. This allows them to be stored for long periods of time and
used at a later date for in vitro fertilization (IVF) or other assisted reproductive technologies.
Cryopreservation can be a useful option for individuals or couples who wish to have children
at a later time, or for those who wish to preserve their fertility due to medical reasons.

Vitrification is a modern cryopreservation technique that has revolutionized the field of

assisted reproduction. It is a method of rapidly freezing eggs, embryos, and sperm at
extremely low temperatures, in order to preserve their integrity and potential for fertilization.
Vitrification is considered a significant improvement over traditional slow-freezing methods,
as it reduces the formation of ice crystals, which can damage the cells being frozen. This
makes vitrification an efficient and effective way to preserve fertility for individuals and
couples who wish to have children at a later time, for those who need to preserve their
fertility due to medical reasons, or for research and conservation purposes. This chapter will
provide a comprehensive overview of the science behind vitrification, its benefits and
limitations, and its current applications in assisted reproduction.
 Having reached a bottleneck in ART in terms of success rates, the endeavour to improve the
pregnancy and livebirth rates has intensified. Whether it is the transfer of a single genetically
normal embryo deemed disease free and euploid by preimplantation genetic testing, the
transfer of embryo/s in a more physiological uterine environment as a part of “freeze-all” and
“segmentation” strategy or simply achieving a higher cumulative success rate by freeze-thaw-
transfer of supernumerary embryos produced by a single stimulation, the cornerstone of all
these modalities is vitrification.

Vitrification and slow-freezing are two methods used in assisted reproduction for the
preservation of oocytes and embryos. Vitrification is a relatively newer technique that
involves freezing the oocyte or embryo at a very fast rate, typically in a matter of seconds.
This rapid cooling rate helps to minimize the formation of ice crystals, which can damage the
cell. As a result, vitrification has been found to have higher survival and pregnancy rates than
slow-freezing. Slow-freezing, on the other hand, involves cooling the oocyte or embryo at a
slower rate, typically over a period of several minutes. This method has been used for many
years and is still used in many clinics. However, it is less efficient than vitrification, and
research has shown that it results in lower survival and pregnancy rates. Ultimately, the
choice between vitrification and slow-freezing will depend on the clinic's equipment and
expertise, and the patient's specific needs and preferences.

There are several indications for using vitrification in assisted reproduction,

including:

1. Oocyte banking: Vitrification allows for the preservation of oocytes for future
use, allowing women to delay childbearing or to preserve fertility before
undergoing treatments that may damage their fertility, such as chemotherapy
or radiation therapy.
2. Embryo banking: Vitrification also allows for the preservation of embryos for
future use, allowing couples who have undergone in vitro fertilization (IVF) to
have additional children without having to repeat the IVF process.
3. Donor oocyte and embryo banking: Vitrification allows for the preservation of
oocytes and embryos from donors for use in assisted reproduction, increasing
the availability of these materials for people who need them.
4. Genetic testing and preimplantation genetic diagnosis (PGD): Vitrification allows
for the preservation of embryos for genetic testing and PGD, allowing couples
to screen for genetic disorders before implantation.
5. Surrogacy: Vitrification allows for the preservation of embryos for use in
surrogacy, allowing intended parents to have a biological child even if they are
unable to carry a pregnancy.
6. Fertility preservation for cancer patients: Some cancer treatments can damage
the ovaries or testicles, making it difficult for the patient to have biological
 children in the future. Vitrification of oocytes or sperm can help preserve the
patient's fertility before undergoing cancer treatments.
7. Cryopreservation for research: Vitrification allows for the preservation of
oocytes and embryos for research, increasing the availability of these materials
for scientists studying reproductive biology.