Fig. 1.

Representative figures of human granulosa cells (GCs) grown in 2% FCS containing medium supplemented with EGF, bFGF and FSH. a GCs was initiated by the addition of folicular fluid into the cultivation medium (day 1). The follicular fluid created a protein surface on the botton of the cultivation flask that facilitated cell adhesion. b Morphology of GCs cultivated without follicular fluid (day 1). c Figure shows a small, rapidly dividing GCs with high proliferative potential, which were initially cultivated with follicular fluid (day 10). d Culture of GCs cultivated without follicular fluid and undergoing the cell degradation process, containing fewer dividing cells (day 10). e Typical spindle-shaped morphology of human GCs initially cultivated with follicular fluid (day 31). f Spindle-shaped cells were not morphologically changed during long-term cultivation; granulosa cells (GC8 line) initially cultivated in media with follicular fluid (day 99). Scale bar 50 μm. g Immunocytochemical analysis of GCs for FSHR (day 69) and h LHR after initial cultivation with FF (day 69). Cells stained with the omission of a primary antibody were used as negative controls. Scale bar 100 μm