function; these ARTs include in vitro growth and maturation of
ovarian follicles followed by in vitro fertilization, intracytoplasmic
sperm injection enabling embryo production, and transfer in to
surrogate mothers [30, 31].
Heterotopic OT grafts are highly depended on the determination of optimal anatomical site which would provide angiogenic
advantage for rapid vascularization. Furthermore, the choice of site
must also provide convenient access for monitoring or follicle
isolation when needed and also has to be transferable to human
reproductive medicine. To this end, various sites (e.g., subcutaneous, omentum, renal capsule, neck muscle, intraperitoneal, and
abdominal wall) have been experimented on rodents [32–
37]. Among these sites, subcutaneous transplantation requires
minimal technical effort and may allow monitoring follicular development using currently available imaging techniques such as ultrasound. In addition, it would be possible to retrieve oocytes from
the grafts, fertilize them in vitro, and transfer the resulting embryos
back to the patient who received the OT autograft to restore
fertility.
The safety, effectiveness, and longevity of cryopreserved OT for
maintaining the endocrine and gametogenic functions after transplantation are important determinants. However, these parameters
are difficult to assess directly in humans due to ethical and safety
problems associated with these potential reproductive therapeutic
approaches. For instance, one of the major concerns is the risk of
transplanting malignant cells, which could be in the grafted
OT. Thus, experimental rodent studies can be designed to address
this concern and obtain valuable information to determine safety of
OT transplantation as an option for human patients with gonadal
dysfunction. In addition, OT cryopreservation for future transplantation were used to investigate menopause or age-related metabolic
and cognitive disorders [25, 38–40]. Rodents as model animals are
invaluable to distinguish the impact of reproductive function or
reproductive aging from natural aging process. In this chapter, we
provide methodologies for rodent OT cryopreservation and potential transplantation strategies.
2 Materials
1. Programmable freezer.
2. Liquid nitrogen, cryostorage tank with sample storage system,
and 2 L capacity dewar flasks.
3. Cryoprotective gear: cryo-apron, cryo gloves, and unvented
safety goggles.
Rodent Ovarian Tissue Cryopreservation
471
ovarian follicles followed by in vitro fertilization, intracytoplasmic
sperm injection enabling embryo production, and transfer in to
surrogate mothers [30, 31].
Heterotopic OT grafts are highly depended on the determination of optimal anatomical site which would provide angiogenic
advantage for rapid vascularization. Furthermore, the choice of site
must also provide convenient access for monitoring or follicle
isolation when needed and also has to be transferable to human
reproductive medicine. To this end, various sites (e.g., subcutaneous, omentum, renal capsule, neck muscle, intraperitoneal, and
abdominal wall) have been experimented on rodents [32–
37]. Among these sites, subcutaneous transplantation requires
minimal technical effort and may allow monitoring follicular development using currently available imaging techniques such as ultrasound. In addition, it would be possible to retrieve oocytes from
the grafts, fertilize them in vitro, and transfer the resulting embryos
back to the patient who received the OT autograft to restore
fertility.
The safety, effectiveness, and longevity of cryopreserved OT for
maintaining the endocrine and gametogenic functions after transplantation are important determinants. However, these parameters
are difficult to assess directly in humans due to ethical and safety
problems associated with these potential reproductive therapeutic
approaches. For instance, one of the major concerns is the risk of
transplanting malignant cells, which could be in the grafted
OT. Thus, experimental rodent studies can be designed to address
this concern and obtain valuable information to determine safety of
OT transplantation as an option for human patients with gonadal
dysfunction. In addition, OT cryopreservation for future transplantation were used to investigate menopause or age-related metabolic
and cognitive disorders [25, 38–40]. Rodents as model animals are
invaluable to distinguish the impact of reproductive function or
reproductive aging from natural aging process. In this chapter, we
provide methodologies for rodent OT cryopreservation and potential transplantation strategies.
2 Materials
1. Programmable freezer.
2. Liquid nitrogen, cryostorage tank with sample storage system,
and 2 L capacity dewar flasks.
3. Cryoprotective gear: cryo-apron, cryo gloves, and unvented
safety goggles.
Rodent Ovarian Tissue Cryopreservation
471