Functional Testing of Primitive Oocyte-like Cells Developed in Ovarian Surface Epithelium Cell Culture from Small VSEL-like Stem Cells: Can They Be Fertilized One Day?

Abstract

Data from the literature show that there are different populations of stem cells present in human adult ovaries, including small stem cells resembling very small embryonic-like stem cells (VSELs). These small ovarian stem cells with diameters of up to 5μm are present in the ovarian surface epithelium and can grow into bigger, primitive oocyte-like cells that express several markers of a germinal lineage and exhibit pluripotency but not the zona pellucida structure when cultured in vitro. In this report, we present the results of the functional testing of such primitive oocyte-like cells from one patient with premature ovarian failure after insemination with her partners' sperm. Knowing that even immature oocytes collected in an in vitro fertilization program cannot be fertilized naturally, we were only interested in determining whether and how these cells react to added sperm and whether spermatozoa somehow "recognize" them. Interestingly, the primitive oocyte-like cells quickly released a zona pellucida-like structure in the presence of sperm. Two different populations of cells were distinguished, those with a thick and those with a thin zona pellucida-like structure. The primitive oocyte-like cells with a released zona pellucida-like structure expressed the pluripotency-related gene OCT4A (POU5F1) and zona pellucida-related gene ZP3, similar to oocytes obtained from in vitro fertilization but not somatic chondrocytes. In a small proportion of these cells, a single-spermatozoon was observed inside the cytoplasm, but no signs of fertilization were found. These observations may suggest a primitive "cortical reaction". Our data further confirm the presence of germinal stem cells in the ovarian surface epithelium cell culture.