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

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.

Follicular factors determining the developmental competence of porcine oocyte.

PURPOSE: This study aimed to examine the relationship between granulosa cells (GCs), number of follicles, and the ability of follicular fluid to support in vitro growth of oocytes. METHODS: The culture medium was supplemented with follicular fluid (FF) collected from GC-rich ovaries and GC-poor ovaries, and its effect on in vitro growth and quality of oocytes derived from early antral follicles (EAFs) was assessed. RESULTS: GC-rich FF treatment enhanced oocyte growth and augmented changes in the chromatin configuration and lipid content of oocytes when compared to oocytes treated with GC-poor FF. Moreover, oocytes treated with GC-rich FF had a higher ability to progress to the blastocyst stage, than oocytes derived from large antral follicles (3-5 mm in diameter). In addition, supplementation of the culture medium with either GC-rich or GC-poor FF enhanced histone acetylation in oocytes grown in vitro. CONCLUSION: GC-rich FF contains key factors that support in vitro oocyte growth; hence, oocytes grown in GC-rich FF medium had high developmental competence, which was comparative to the oocytes grown in vivo.

Modification of the medium volume and gel substrate under in vitro culture conditions improves growth of porcine oocytes derived from early antral follicles.

This study compared the effects of different volumes of culture medium for the in vitro growth of oocytes derived from porcine early antral follicles (EAFs). Oocyte granulosa cell complexes (OGCs) were collected from EAFs (0.5-0.7 mm in diameter) and individually cultured for 14 days. When OGCs were cultured in 1 ml of medium with or without polyacrylamide gel (PAG), the presence of PAG supported granulosa cell (GC) proliferation and oocyte growth. When OGCs were cultured in 0.2 or 1 ml of medium on PAG, the number of GC in the OGC culture and the developmental ability of the oocytes cultured in vitro were significantly higher for the 1 ml of culture medium group than for the 0.2 ml group. In conclusion, a combination of a large volume of culture medium with PAG improved the growth and developmental ability of the oocytes cultured in vitro, which were comparable to the oocytes collected from large antral follicles.