For girls and women, therapeutic cancer treatments such as chemo or radiation therapy can have a harmful affect on the germ cells within their reproductive organs. This is a particularly harsh consequence because mammalian females are born with all the eggs they will ever have. If this pool of eggs is destroyed, the result is sterility. Although the technology does not yet exist for humans, some women and young girls about to undergo these treatments are currently having portions of their ovaries excised and cryogenically frozen in hopes that a process that can restore fertility is developed. Since growing follicles do not survive the freezing process, only the small primordial follicles remain viable after cryopreservation. Therefore, the focus of my research is to create an in vitro culture system that can support the growth and maturation of primordial ovarian follicles in order for these women to bear their own genetic offspring.

Anatomy of a Rat Preantral Ovarian Follicle.

Immobilization methods for the fabrication of amperometric biosensors often rely on the entrapment of enzyme in a gel layer, which may be further coated by an external protective film. Interestingly, in the case of polyurethane/hydrophobic polyurethane based biosensors, the enzyme may be directly dispersed in the external hydrophilic polyurethane film. The lifetime and use of such systems are often limited by the diffusion of enzyme through the external membrane. To overcome this main disadvantage, the enzyme has to be directly and covalently immobilized into the coating.

In the past, the conventional technique has attempted to grow these follicles on a flat surface in static medium. Although this environment has been successful in maturing mouse follicles, the development of follicles from larger species, such as rats, pigs, and humans, with this system has failed. Our preliminary studies on rat follicles have found that under these conventional conditions, rat follicles experience flattening and rupture with a loss of anatomic integrity. However, when rat follicles were cultured orbiting test tube - suspension culture system and rotating wall vessels, the follicles did not rupture and maintained the spherical geometry associated with healthy follicles.

Therefore, our current working hypothesis is that the static conventional follicle culture systems are unable to support follicle development beyond the antral transition because of inadequate metabolic support, inadequate architectural support, or inadequacies in both.