Figure 5.

PMSG-primed primary granulosa cells differentiate in culture. (A) Primary granulosa cells secrete estradiol and P₄ throughout the 7-day culture period. The levels of E₂ and P₄ in the culture medium steadily increase throughout the culture period. (B) The increase in hormone production is consistent with increasing expression of Star, Cyp19a1, and Cyp11a1. (C) Primary granulosa cells are proliferative in culture as identified using an EdU incorporation assay. This proliferative potential diminishes as the culture progresses, and cells eventually become contact inhibited (data not shown). n = 3; *P < 0.05 compared with 1-day culture unless otherwise indicated.