Effects of isolating methods (mechanical or enzymatical) on structure of pre-antral follicles in mouse.

PURPOSE: This study was conducted to evaluate the stable and effective isolating method of pre-antral follicles from ovary by means of mechanical or enzymatical method in mice. METHODS: Mouse (ICR, 3-4 weeks old) ovarian pre-antral follicles of 70-130 microm (A: < or = 90; B: 91-110; C: 111-130 microm) in diameter were isolated by mechanical method (group I) using syringe needles or by enzymatical method (group II) using 600 IU collagenase and DNase and cultured individually in 20 microL droplet of media under mineral oil on culture dish for 8 days. RESULTS: On 8th day after culture, disruption rate (84.4 +/- 18.8% vs. 9.4 +/- 18.8%) was significantly higher (p < 0.05) in group II than in group I. Survival rate (88.0 +/- 28.9% vs. 0%) was significantly higher (p < 0.0000) in group I than in group II. There were no differences in the survival rate among different initial sized pre-antral follicles in group I (A: 86.7 +/- 12.5%, B: 100%, C: 69.2 +/- 30.0%, NS) or group II (A: 0%, B: 0%, C: 0%, NS), respectively. CONCLUSIONS: In conclusion, compared to mechanical dissection, enzymatical isolation of pre-antral follicles resulted in higher follicular disruption rate. Survival rate was not affected by different initial sizes of pre-antral follicles in mice.

The expression of matrix metalloproteinase-9 in human follicular fluid is associated with in vitro fertilisation pregnancy.

OBJECTIVE: To investigate the role of matrix metalloproteinase-9 (MMP-9) in the pre-ovulatory follicular fluid and culture media during in vitro fertilisation (IVF) cycle and to develop the zymographic pre-diagnosis marker for successful implantation and pregnancy in human IVF. DESIGN: Controlled clinical study. SETTING: IVF Laboratory, Women's Hospital Infertility Clinic and Dongguk University, Korea. SAMPLE: Women undergoing in vitro fertilisation treatment. METHODS: Experiments were designed for controlled clinical study with women undergoing IVF treatment. MMP-9 expressions in follicular fluid and culture media samples that had been collected during transvaginal oocyte retrieval were measured using zymography. MMP-9 activities and expressions were strongly correlated to a higher rate of fertilisation and pregnancy. MAIN OUTCOME MEASURES: Fertilisation rates and ultrasonic evidence of intrauterine pregnancy by four weeks after embryo transfer. RESULT: MMP-9 activity was significantly higher in the pregnant group than in the non-pregnant group (P < 0.01). In contrast, MMP-2 activity was present in the follicular fluid and culture media of all women, and no difference in its expressions was found between the pregnant and non-pregnant groups. No correlation was found between the MMP-9 expression in follicular fluid and culture media and the fertilisation rates. CONCLUSION: The expression of MMP-9 in the follicular fluid and culture media is a prerequisite for successful pregnancy in IVF cycle. The zymography of MMP-9 activity in follicular fluids of human and culture media was developed as a pre-diagnostic method and zymographic diagnosis marker for successful fertilisation, implantation and pregnancy in human IVF.