Figure 4
From: Oogenesis in cultures derived from adult human ovaries
Development of oocytes in day 3 to 5 mixed ovarian cultures. Undifferentiated (stem) cells persist in all day 3 cultures (a). Under PhR+ conditions (b-h), the 100 μm oocyte-like cells were found on day 5 (arrows, b) with developing zona pellucida layer (arrow, c) in the presence of associated fibroblasts (arrowheads). Association of granulosa type cells (arrowheads, d) appears to stimulate advanced oocyte growth. Thick zona pellucida layer is apparent in some oocytes accompanied by fibroblasts on day 4 (e), and staining for ZP proteins on day 5 shows no change in the oocyte size but a lack of surface ZP expression (arrow, f). Staining for CK5,6,8,17 (g) shows cytoplasmic expression in mesenchymal type cells (black arrowhead) and non-specific staining (peroxidase expression) in the nuclei of other cells (arrowheads). h) Isolated large (200 μm) cells showed characteristics of secondary oocytes, including poor separation between the nucleus and cytoplasm (black arrowhead) expulsion of polar body (white arrowhead) and surface expression of ZP proteins (black arrow). The latter was, however, not apparent in the surface segment close to the polar body (white arrow). Under PhR- conditions, rare giant oocytes (300 μm) were found on day 4 (i). They lacked the germinal vesicle breakdown and surface expression of ZP proteins on day 5 (arrow, j). Bar in (i) for a-f and h-j.