O-16: Comparison of Pre-Antral Follicle Culture Development during 2 Dimensional and 3 Dimensional Culture Systems

Background: Setting up an in vitro follicle culture system that resembles in vivo ovary condition has high value in research. Additionally, expression evaluation of folliculogenesis involved genes could lead us to the designing of better culture system. Materials and Methods: ovaries of 12-day-old female NMRI mice were removed, 100-130 μm pre-antral follicles were mechanically isolated from fresh ovaries and cultured for 12 days in two-dimensional (2D) and threedimensional (3D) culture systems. Alginate hydrogel (0.8%) was chose as a matrix for 3D culture and α-MEM medium that supplemented with FSH, LH, ITS, Activin and FBS was consisted as a Culture medium. Follicles survival rate, growth and also quantitative expression of oocyte maturation genes (Gdf9, Bmp15, Bmp6) were analyzed after 24 hours and 12 days of culture. Results: In 2D culture system, a higher survival rate was observed in comparison with 3D culture system (p<0.05). During culture period, significant follicle diameter increase was observed. oocyte maturation genes relative expression indicated non significant higher expression of three mentioned genes in 3D culture system than 2D system. Conclusion: As 3D culture system maintains oocyte maturation m-RNA reserve better than 2D culture system, it seems that improving of 3D culture condition could cause better follicle survival, development and growth.