Production of Ovarian Cell Types from Human Pluripotent Stem Cells

Abstract

The ovary is the central organ of female reproduction, and understanding it is crucial for improving women’s health and treating infertility. However, studies of human ovarian development are limited by technical and ethical challenges in accessing ovarian tissue, especially during fetal stages. Therefore, engineering stem cell-based models of the human ovary in vitro has the potential to rapidly advance the study of human reproduction by enabling experiments that would not otherwise be possible. Furthermore, if the full process of human oogenesis can be re-created in vitro starting from pluripotent stem cells, this would be a transformative therapy for millions of people around the world who are infertile due to a lack of eggs. Although in vitro oogenesis has recently been achieved in mice, this has not yet been successful for humans, largely due to a lack of methods for producing the necessary ovarian cell types. In this dissertation, I present my work on directing the differentiation of human induced pluripo- tent stem cells (hiPSCs) into ovarian cell types in vitro, including granulosa-like, oogonia-like, and mei- otic cells. My overall approach has centered on using computational tools and multiplexed screens to identify key regulatory factors that drive the formation of a cell type of interest. In Chapter 1, I discuss the production of granulosa-like cells and their application in forming ovarian organoids. In Chapter 2, I showcase a method for the rapid generation of oogonia-like cells with erasure of DNA methy- lation. In Chapter 3, I describe my latest work on directly initiating meiosis from hiPSCs. Overall, these methods establish in vitro models of various aspects of ovarian development that will be useful to researchers in the field, and may serve as the basis for future therapies such as in vitro oogenesis.