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KMID : 0900919990230010053
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Korean journal of Animal Reproduction
1999 Volume.23 No. 1 p.53 ~ p.61
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Effects of Gonadotrophins on In Vitro Growth and Maturation of Mouse Preantral Follicles
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Kim Dong-Hoon
Kang Hee-Gyoo
Han Sung-Won
Lee Hoon-Taek
Lee Ho-Joon
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Abstract
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The present study was designed to investigate the effects of gonadotrophins on in vitro growth and maturation in mouse preantral follicles. Ovaries were removed from 12-day-old ICR mice. Follicles were dissociated enzymetically in Leibovitz L-15 medium containing 1 mg/mell collagenase and 0.2 mg/mell DNase I. The follicles were cultured on Transwell-COL membrane inserts in six well cluster dishes for 10 days. The culture medium was alphaMEM medium supplemented with 5% fetal bovine serum and FSH or HMG. After 10 days of growth in vitro, follicles were allowed to mature for 18~20 hr in medium supplemented with 1.5 IU/mell hCG. The oocytes were then denuded of their cumulus cells and assessed maturation status. Concentrations of oestradiol and progesterone were measured with a radioimmunoassay. Oocyte diameter was determined with an ocular micrometer. The survival and Metaphase II rates of oocytes were significantly higher in FSH treatment groups than in control group (P＜0.001), but there were no differences among the groups of treated FSH concentration. The survival and Metaphase II rates of oocytes in HMG treatment group (60.9 and 40.6%) were higher than in FSH treatment group (76.6 and 48.2%) and control group (49.2 and 7.1%). The survival and Metaphase II rates of oocytes on both FSH and LH treatment groups were no differences among the ratios of FSH and LH. Diameter of oocyte was no differences among the treatment groups, but smaller than compared to in vivo grown oocyte. Through the entire culture period, secretions of oestradiol and progesterone were significantly less in control group than in HMG and FSH treatment groups. These results suggest that gonadotrophins playa key role in in vitro culture of mouse preantral follicles. Especially, addition of FSH and LH should be more effective than FSH alone.
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KEYWORD
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Preantral follicle, In vitro growth, FSH, LH, HMG.
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