Chapter 22
Cryopreservation and Transplantation of Laboratory Rodent
Ovarian Tissue for Genome Banking and Biomedical
Research
Yuksel Agca and Cansu Agca
Abstract
Genetic modifications in combination with highly sophisticated assisted reproductive technologies such as
in vitro oocyte maturation and development, in vitro fertilization, intracytoplasmic sperm injection, and
in vitro embryo culture have opened many research avenues and treatment options for both animals and
humans. The number of genetically modified (GM) rodent strains increased considerably during the last
several decades, and their numbers are expected to increase due to efficient gene editing technologies
including the CRISPR/Cas9. Rodent ovarian tissues (OT) cryopreservation and transplantation procedures have several applications in biomedical field: they provide a fertility restoration option for GM rodent
strains in some circumstances. They also serve as models to investigate OT cryopreservation as potential
alternatives for human infertility patients as well as other domestic and wildlife species for the development
of improved cryopreservation and subsequent transplantation strategies. The modeling studies enable
determining effective cryoprotective agents (CPA), CPA and water permeability kinetics, and cooling and
warming rates during the development of OT cryopreservation procedures. Furthermore, rodent models
are extremely useful for determining post-thaw OT graft sites as well as potential medical interventions in an
effort to expedite angiogenesis and inhibit inflammatory/immune response, OT longevity, and follicular
integrity. Here we describe methodologies for rodent OT cryopreservation and potential transplantation
sites for frozen-thawed rat and mouse OT.
Key words Mouse, Rats, Ovary cryopreservation, Transplantation
1 Introduction
Historically, rodent ovarian tissue (OT) cryopreservation and transplantation studies can be divided into two phases. The earlier
studies were performed in the 1950s, when glycerol was the only
known cryoprotective agents (CPA). Parkes and Smith [1] cryopreserved rat OT by slowly cooling them to À79
C in the presence
of 15% glycerol and were able to obtain partial restoration of
reproductive cycle. A few other studies during the 1950s investigated longevity of transplanted ovaries as well as the incidence of
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_22, © Springer Science+Business Media, LLC, part of Springer Nature 2021
469
Cryopreservation and Transplantation of Laboratory Rodent
Ovarian Tissue for Genome Banking and Biomedical
Research
Yuksel Agca and Cansu Agca
Abstract
Genetic modifications in combination with highly sophisticated assisted reproductive technologies such as
in vitro oocyte maturation and development, in vitro fertilization, intracytoplasmic sperm injection, and
in vitro embryo culture have opened many research avenues and treatment options for both animals and
humans. The number of genetically modified (GM) rodent strains increased considerably during the last
several decades, and their numbers are expected to increase due to efficient gene editing technologies
including the CRISPR/Cas9. Rodent ovarian tissues (OT) cryopreservation and transplantation procedures have several applications in biomedical field: they provide a fertility restoration option for GM rodent
strains in some circumstances. They also serve as models to investigate OT cryopreservation as potential
alternatives for human infertility patients as well as other domestic and wildlife species for the development
of improved cryopreservation and subsequent transplantation strategies. The modeling studies enable
determining effective cryoprotective agents (CPA), CPA and water permeability kinetics, and cooling and
warming rates during the development of OT cryopreservation procedures. Furthermore, rodent models
are extremely useful for determining post-thaw OT graft sites as well as potential medical interventions in an
effort to expedite angiogenesis and inhibit inflammatory/immune response, OT longevity, and follicular
integrity. Here we describe methodologies for rodent OT cryopreservation and potential transplantation
sites for frozen-thawed rat and mouse OT.
Key words Mouse, Rats, Ovary cryopreservation, Transplantation
1 Introduction
Historically, rodent ovarian tissue (OT) cryopreservation and transplantation studies can be divided into two phases. The earlier
studies were performed in the 1950s, when glycerol was the only
known cryoprotective agents (CPA). Parkes and Smith [1] cryopreserved rat OT by slowly cooling them to À79
C in the presence
of 15% glycerol and were able to obtain partial restoration of
reproductive cycle. A few other studies during the 1950s investigated longevity of transplanted ovaries as well as the incidence of
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_22, © Springer Science+Business Media, LLC, part of Springer Nature 2021
469