Single-cell RNA sequencing analysis of porcine ovarian somatic cells in antral follicles

The antral follicle stage plays a crucial role in mammalian oocyte maturation, signifying the final stages of oocyte development and ovulation. This complex process relies on synchronized interactions between oocyte maturation and the proliferation of neighboring granulosa cells. Previous studies have identified two subtypes of granulosa cells in antral follicles: cumulus granulosa cells, located in the inner region, which surround and support the oocyte, and mural granulosa cells, present in the outer layers, which provide mechanical support to the follicular wall and possess steroidogenic functions. Despite the wealth of molecular data generated by these studies, many fundamental questions regarding key developmental events, granulosa cell heterogeneity, functional annotation, and the intricate relationship between somatic cells and oocytes still lack detailed, single-cell resolution-level investigations. In this study, we isolated follicular cells from porcine antral follicles and conducted scRNA-seq to analyze the single-cell transcriptomes of these cells. By identifying and sub-clustering ovarian cells, such as mural granulosa cells and cumulus granulosa cells, we elucidated the heterogeneity of granulosa cells in pigs. In this study, we isolated follicular cells from two healthy porcine antral follicles and conducted scRNA-seq using 10X genomics platform to analyze the single-cell transcriptomes of these cells.