Oncofertility Publications

Abstract

In vitro ovarian follicle cultures may provide fertility-preserving options to women facing premature infertility due to cancer therapies. An encapsulated three-dimensional (3-D) culture system utilizing biomaterials to maintain cell-cell communication and support follicle development to produce a mature oocyte has been developed for the mouse. We tested whether this encapsulated 3-D system would also support development of nonhuman primate preantral follicles, for which in vitro growth has not been reported. Three questions were investigated: Does the cycle stage at which the follicles are isolated affect follicle development? Does the rigidity of the hydrogel influence follicle survival and growth? Do follicles require luteinizing hormone (LH), in addition to follicle-stimulating hormone (FSH), for steroidogenesis? Secondary follicles were isolated from adult rhesus monkeys, encapsulated within alginate hydrogels, and cultured individually for </=30 days. Follicles isolated from the follicular phase of the menstrual cycle had a higher survival rate (P < 0.05) than those isolated from the luteal phase; however, this difference may also be attributed to differing sizes of follicles isolated during the different stages. Follicles survived and grew in two hydrogel conditions (0.5% and 0.25% alginate). Follicle diameters increased to a greater extent (P < 0.05) in the presence of FSH alone than in FSH plus LH. Regardless of gonadotropin treatment, follicles produced estradiol, androstenedione, and progesterone by 14-30 days in vitro. Thus, an alginate hydrogel maintains the 3-D structure of individual secondary macaque follicles, permits follicle growth, and supports steroidogenesis for </=30 days in vitro. This study documents the first use of the alginate system to maintain primate tissue architecture, and findings suggest that encapsulated 3-D culture will be successful in supporting the in vitro development of human follicles.

Min Xu, Erin R. West-Farrell, Richard L. Stouffer, Lonnie D. Shea, Teresa K. Woodruff, and Mary B. Zelinski; Biology of Reproduction(3):587-94 Sep.8, 2009

Abstract

OBJECTIVE:
To investigate the effect of slow cryopreservation on the morphology and function of primate primordial follicles within ovarian tissue slices.

DESIGN:
Fresh monkey ovarian tissue was frozen by slow cryopreservation and thawed for analysis of morphologic and functional parameters.

SETTING:
University-affiliated laboratory.

ANIMALS:
Rhesus monkey ovarian tissue.

INTERVENTION(S):
None.

MAIN OUTCOME MEASURE(S):
Histologic analysis, follicle counting, assessment of protein abundance and localization.

RESULT(S):
After freezing and thawing, 89% of the primordial follicles maintained their laminar-based architecture, with sizes close to those of fresh fixed follicles. Molecular markers of early follicle health (activin subunits and the phosphorylated form of the signaling protein Smad2 [pSmad2]) were present in fresh and frozen-thawed primordial follicles. Stroma cells, but not follicles, had a higher level of TUNEL staining. Granulosa cells within the follicles of frozen-thawed ovarian tissue cultured for 48 hours had the capacity to proliferate and sustained expression of the activin subunits and nuclear pSmad2.

CONCLUSION(S):
This study provides evidence that markers of early follicle growth and development are preserved after slow cryopreservation and thaw, with little effect on follicle morphology and function.

Shiying Jin, PhD, Lei Lei, PhD, Lonnie D. Shea, PhD, Mary B. Zelinski, PhD, Richard L. Stouffer, PhD, and Teresa K. Woodruff, PhD. Fertility and Sterility, 2010.

Abstract

Children and women of reproductive age are increasingly surviving cancer diagnoses, and therefore long-term quality-of-life issues are of greater importance at the time of diagnosis. Cancer therapies including radiation and chemotherapy can be detrimental to fertility, and therefore many patients are motivated to preserve fertility prior to cancer treatment. The only highly successful method in preserving fertility to date is embryo cryopreservation, which may not be appropriate for some patients due to age, delay in treatment, cancer type and stage, as well as availability of an acceptable sperm donor. Alternative methods including oocyte cryopreservation and ovarian tissue banking may also preserve fertility while providing additional flexibility to patients. In vitro ovarian follicle maturation following tissue banking is one potential approach that would not require a delay in cancer therapy for ovarian stimulation, would not require an immediate sperm donor, and does not carry the risk of reintroducing malignant cells following tissue transplantation. In vitro follicle culture systems have resulted in successful live births in the mouse. However, many challenges must be addressed in translating the system to the human. This review summarizes current approaches to fertility preservation and discusses recent developments and future challenges in developing a human in vitro follicle culture system.

Erin R. West, PhD, Mary B. Zelinski, PhD, Laxmi A. Kondapalli, MD, Clarisa Gracia, MD, Jeffrey Chang, MD, Christos Coutifaris, MD, PhD, John Critser, PhD, Richard L. Stouffer, PhD, Lonnie D. Shea, PhD, and Teresa K. Woodruff, PhD; Pediatr Blood Cancer 2009;53:289–295