Identification and characterization of epithelial cells derived from XXXX ovarian follicular fluid.

@entity1 @entity17938 @entity24262 @entity4729 @entity2440 @entity28374 @entity44 @entity19 @entity30137 @entity28866 @entity3893
INTRODUCTION: Follicular fluid is important for follicular development and oocyte maturation. Evidence suggests that follicular fluid is not only rich in proteins but cells. Besides oocytes, the follicular fluid contains granulosa, thecal, and ovarian surface epithelial cells, and both granulosa and thecal cells are well-characterized. However, epithelial cells in follicular fluid are poorly studied. This study aims to isolate and characterize in vitro epithelial cells that originate from @entity1 ovarian follicular fluid retrieved in the assisted fertilization program. METHODS: Follicular fluid samples were collected from 20 @entity1 in the assisted reproduction program. Epithelial cells were characterized by flow cytometry assay, immunofluorescence staining, real-time PCR, and time lapse photography. RESULTS: Epithelial cell cultures were established from 18 samples. A small population of epithelial cells expresses germ-line stem cell markers, such as @entity4729 ( @entity4729 ), @entity2440 , and @entity30137 ( @entity30137 ). In the epithelial cell culture system, oocyte-like cells formed spontaneously in vitro and expressed the following transcription markers: deleted in @entity3893 -like ( @entity28866 ), developmental pluripotency associated protein 3 @entity28374 -related protein ( @entity28374 ), zona pellucida gene family C (ZPC), @entity17938 ( @entity17938 ), and @entity24262 ( @entity24262 ). Some of the oocyte-like cells developed a zona pellucida-like structure. Both the symmetric and asymmetric division split of epithelial cells and early developing oocytes were observed using time lapse photography. Cell colonies were formed during epithelial culturing, which maintained and proliferated in an undifferentiated way on the feeder layer and expressed some pluripotency markers. These colonies differentiated in vitro into various somatic cell types in all three germ layers, but did not form @entity44 when injected into immunodeficient @entity19 . Furthermore, these epithelial cells could be differentiated directly to functional hepatocyte-like cells, which do not exist in ovarian tissues. CONCLUSIONS: The epithelial cells derived from follicular fluid are a potential stem cell source with a pluripotent/multipotent character for safe application in oogenesis and regenerative medicine.
