Establishment female germ line stem cell from pig ovary and investigate differentiation potential

Abstract

According to classical knowledge of reproductive biology, there was a limited number of oocytes and no possibility of renewal if the oocytes were lost due to disease and injury in the ovary of female mammalians. However, recent studies suggested that female germ line stem cells (FGSCs) replenished oocyte-pools in adult stage. Hence, this study was to explore the ideal medium for successfully establishment FGSCs in ovarian surface epithelium (OSE) in porcine ovaries and differentiation them into oocyte-like cells (OLCs).Here, FGSCs was culture in three different medium including DMEM /F12 supplied with 10% FBS, DMEM/ F12 supplied with 10% FBS and growth factors, DMEM/F12 supplied with N2l, growth e and invetigate differentiation capabilities of FGSCs when aggregation with granulosa cells as monolayer. The result showed that FGSCs were small spherical morphology, dark, shiny with a small diameter from 10-15um and fully grew under suitable medium as free-serum DMEM-F12 supplied with N21 and growth factors. The expression of germline stem cell markers as Vasa were analysed by immunofluorescence staining. Our results also demonstrated that granulosa cells (GCs) were effective in inducing the differentiation of FGSCs into OLCs through direct cell-to-cell contacts. As a result, this study revealed that when culturing FGSCs under free-serum medium supplied with N2l and growth factors, the development of population FGSCs was enhanced and the FGSCs communicated with GCs had developmental potentials including spontaneous differentiation into oocyte-like cells. Keywords: Female germ line stem cells, Oocyte-like cells, Ovarian surface epithelium, Granulosa cells