In vitro differentiation of human oocyte-like cells from oogonial stem cells: single-cell isolation and molecular characterization

AbstractSTUDY QUESTIONAre the large cells derived from cultured DEAD box polypeptide 4 (DDX4)-positive oogonial stem cells (OSCs), isolated from the ovarian cortex of non-menopausal and menopausal women, oocyte-like cells?SUMMARY ANSWERUnder appropriate culture conditions, DDX4-positive OSCs from non-menopausal and menopausal women differentiate into large haploid oocyte-like cells expressing the major oocyte markers growth differentiation factor 9(GDF-9) and synaptonemal complex protein 3 (SYCP3) and then enter meiosis.WHAT IS KNOWN ALREADYThe recent reports of OSCs in the ovaries of non-menopausal and menopausal women suggest that neo-oogenesis is inducible during ovarian senescence. However, several questions remain regarding the isolation of these cells, their spontaneous maturationin vitro, and the final differentiation state of the resulting putative oocytes.STUDY DESIGN, SIZE, DURATIONDDX4-positive OSCs were obtained from 19 menopausal and 13 non-menopausal women (who underwent hysterectomy for uterine fibroma, ovarian cyst, or other benign pathologies) and cultured for up to 3 weeks. Large and small cells were individually isolated and typed for early and late differentiation markers.PARTICIPANTS/MATERIALS, SETTING, METHODSOvarian cortex fragments were processed by immuno-magnetic separation using a rabbit anti-human DDX4 antibody and the positive populations were measured by assessing both FRAGILIS and stage-specific embryonic antigen 4 (SSEA-4) expression. After 3 w...
Source: Human Reproduction - Category: Reproduction Medicine Source Type: research