Effect of exogenous PLAU and SERPINE2 on matrix gene expression and the hyaluronan status of cumulus cells during IVM.

COCs were incubated with SERPINE2 (0.06 mg/ml) or PLAU (20 IU) in IVM medium for 3, 6, or 16 h. After culturing, cumulus cells were collected for qRT-PCR analyses or COCs were transferred onto slides and fixed for hyaluronan evaluation. A, qRT-PCR revealed Has2 (a), Vcan (b), Tnfaip6, and Ptx3 mRNA levels in cumulus cells. Bars indicate means ± SD of three independent experiments each. P<0.05, *P<0.0001 compared with the group that COCs cultured for 3 h in IVM medium. B, After culturing for 6 h, the hyaluronan contents in untreated COCs (a and e) or COCs treated with PLAU (c and g) or SERPINE2 (d and h) were revealed by treatment with or without (staining control, b and f) HABP (green) as described in Materials and Methods. For contrast, the slides were counterstained with Hoechst 33258 (blue, a–d) or photographed under differential interference contrast microscopy (e–h). Scale bars, 100 µm.