Cell-free DNA content in follicular fluid: A marker for the developmental ability of porcine oocytes.

ABSTRACT

PURPOSE: The present study examined the relationships among the amount of cell-free-DNA (cfDNA) in porcine follicular fluid (FF), the developmental ability of enclosed oocytes, and characteristics of granulosa cells and examined the effect of cfDNA content in maturation medium on the developmental ability of the oocytes. METHODS: Oocytes and FF were collected from individual gilts, and the gilts were rated based on the ability of their oocytes to develop to the blastocyst stage and the amount of cfDNA in the FF. The copy numbers of mitochondrial DNA (Mt-DNA) and nuclear DNA (N-DNA) were measured by real-time PCR and the DNA sequence. FF or cfDNA was added to the maturation medium, and the developmental ability of the oocytes was examined. RESULTS: The amount of cfDNA was associated with apoptosis of the granulosa cells, and high-cfDNA content in FF was associated with low developmental ability of oocytes. Supplementation of the maturation medium with FF containing high cf-Mt-DNA or with DNA extracted from the FF did not affect oocyte developmental competence. CONCLUSIONS: Cell-free DNA content in FF is a marker for oocyte competence, but cfDNA in the oocyte maturation environment did not affect oocyte developmental ability.