Fig. 4.

Blastocysts have significantly more cells after 24 h of culture, but BR-DIM or metformin significantly decreases cell growth compared with unstimulated embryos. Morulae were cultured overnight to acclimate to culture to blastocyst stage, one group was fixed and counted for cells and the rest were preloaded with CC (5 μM) for 2 h and then continued +/− CC with stimuli BR-DIM (20 μM) or metformin (40 μM), for an additional 24 h, and then fixed cells were counted and graphed. Triplicate biological experiments were performed using 174 embryos, tested for normal distribution by ANOVA and significance by Dunnett’s post hoc t test. Significance compared with KSOM (a) and lack of significance for each stimulus with CC compared with the stimulus alone (b)