Figure 9 A-I. Effect of metformin on progesterone production in cultured granulosa cells.
Granulosa cells from prehierarchical follicles (6–8 mm), the most recently recruited preovulatory follicle (9–12 mm) and the smallest preovulatory follicle (F5/6) were cultured and treated with 0, 1, 10 or 20 mM metformin for 24, 48 or 72 h in the presence or absence of rhFSH (100 ng/ml) at 37°C. Progesterone concentrations in the cell culture media were quantified by enzyme-linked immunosorbent assay, as described in the Materials and Methods section. Progesterone quantity was normalized to the total protein concentrations in the granuloma cell lysate and analyzed. Control received no rhFSH and no metformin treatments. A, BP < 0.05; n=4 replicate incubations of 4 broiler breeder hens each. Inset: Photograph of prehierarchical follicles and the most-recently recruited preovulatory follicle to denote the source of granulosa cell (open box).