RII Track-4: Using the Cryopreservation and Microfluidic Technologies to Engineer Bottlenose Dolphin Ovarian Function for Testing Marine Contaminants' Female Reproductive Toxicity

University Of South Carolina At Columbia, Columbia SC

Investigators

Abstract

Nontechnical description of the project The bottlenose dolphin is the most common marine mammal along the U.S. coastal states and is designated as a federally protected species under the Marine Mammals Protection Act (MMPA). As one of the apex predators and by having extensive lipid-rich blubber layers, the bottlenose dolphin is more susceptible to the bioaccumulation of lipid soluble marine contaminants. Many of these contaminants are endocrine disrupting chemicals (EDCs) such as polychlorinated biphenyls (PCBs) and phthalates, that can adversely impact the female reproductive functions of marine mammals. Studies related to investigating the effect of marine contaminants on female bottlenose dolphin reproduction are dependent on recording whether a dolphin can be pregnant and produce a viable calf. However, it is difficult to monitor the specific impact of EDC's on the ovary and its related functions. The central objective of this research proposal is to use the cryopreservation and 3D microfluidic technologies to engineer the bottlenose dolphin ovarian function in vitro. The proposed research will allow the establishment of a high-throughput ovarian toxicity screening platform to determine the reproductive toxicity of marine contaminants on female bottlenose dolphins as well as other marine mammals. Technical description of the project The ovary is the primary female reproductive organ and contains follicles at various developmental stages. The follicle primarily functions to secrete sex steroid hormones and to support its enclosed germ cell oocyte for maturation, ovulation, and fertilization. The goal of the proposed research is to visit the host site at Oregon Health and Science University to learn ovarian tissue cryopreservation and the in vitro growth of follicles from large mammalian species. The first research objective of research one is to cryopreserve mouse ovarian follicles through vitrification for establishing a long-term follicle storage platform. The PI will culture follicles in vitro and examine the follicle reproductive outcomes to investigate the effect of vitrification on follicle and oocyte health and also determine the vitrification-induced cell injury. The second research objective is to cryopreserve bottlenose dolphin ovarian follicles and use microfluidic technology to engineer dolphin ovarian function in vitro. We will collect and cryopreserve ovaries from fresh-dead, stranded bottlenose dolphins, use the microfluidic platform to support a long-term dolphin follicle development and maturation, and validate this system by testing known ovarian toxic chemicals. The proposed work will allow the development of a high-throughput platform to study the female reproductive toxicity of marine contaminants on female bottlenose dolphins and enhance PI and his team's research ability in female reproductive toxicology in marine animals. The broader impact of this study will result in a platform that can be used for cryopreserving ovarian tissues and for other cell/tissue types. Microfluidic culture will provide a novel method to three-dimensionally grow mammalian cells/tissues in vitro. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.