Figure 1.

Preparation and rating of follicular fluid (FF) based on the developmental ability of oocytes (A) or on cell‐free mitochondrial (cf‐Mt‐) DNA content (B). A, Follicular contents were aspirated from antral follicles (3‐6mm in diameter) of 40 individual gilts. Thirty COCs were randomly selected and subjected to in vitro maturation and development following activation. The 40 gilts were rated based on the developmental rate of the corresponding oocytes to the blastocyst stage and were divided into 5 groups (from highest to lowest, with each group containing 8 gilts). FF from the highest and lowest groups was used to create High‐FF and Low‐FF. To obtain enough volume of FF for experiments, 2 randomly selected FFs were equally mixed, and 4 lots of FFs were prepared. B, FFs were collected from 20 gilts, and cf‐Mt‐DNA content in the FF was determined by real‐time PCR. The FFs were rated based on cf‐Mt‐DNA content and divided into 5 groups (from highest to lowest, each group containing 4 gilts). The top 4 and bottom 4 FFs were equally mixed to generate High‐cfDNA‐FF and Low‐cfDNA‐FFs. Six lots of High‐ and Low‐cfDNA‐FFs were prepared using a total of 120 gilts